William H. Robinson, MD PhD

All Publications

• Clonally Expanded B Cells in Multiple Sclerosis Bind EBV EBNA1 and GlialCAM. Nature Lanz, T. V., Brewer, R. C., Ho, P. P., Moon, J. S., Jude, K. M., Fernandez, D., Fernandes, R. A., Gomez, A. M., Nadj, G. S., Bartley, C. M., Schubert, R. D., Hawes, I. A., Vazquez, S. E., Iyer, M., Zuchero, J. B., Teegen, B., Dunn, J. E., Lock, C. B., Kipp, L. B., Cotham, V. C., Ueberheide, B. M., Aftab, B. T., Anderson, M. S., DeRisi, J. L., Wilson, M. R., Bashford-Rogers, R. J., Platten, M., Garcia, K. C., Steinman, L., Robinson, W. H. 2022

  Abstract

  Multiple sclerosis (MS) is a heterogenous autoimmune disease in which autoreactive lymphocytes attack the myelin sheath of the central nervous system (CNS). B lymphocytes in the cerebrospinal fluid (CSF) of MS patients contribute to inflammation and secrete oligoclonal immunoglobulins1,2. Epstein-Barr virus (EBV) infection has been linked to MS epidemiologically, but its pathological role remains unclear3. Here we demonstrate high-affinity molecular mimicry between the EBV transcription factor EBNA1 and the CNS protein GlialCAM, and provide structural and in-vivo functional evidence for its relevance. A cross-reactive CSF-derived antibody was initially identified by single-cell sequencing of the paired-chain B cell repertoire of MS blood and CSF, followed by protein microarray-based testing of recombinantly expressed CSF-derived antibodies against MS-associated viruses. Sequence analysis, affinity measurements, and the crystal structure of the EBNA1-peptide epitope in complex with the autoreactive Fab fragment allowed for tracking the development of the naïve EBNA1-restricted antibody to a mature EBNA1/GlialCAM cross-reactive antibody. Molecular mimicry is facilitated by a post-translational modification of GlialCAM. EBNA1 immunization exacerbates the mouse model of MS and anti-EBNA1/GlialCAM antibodies are prevalent in MS patients. Our results provide a mechanistic link for the association between MS and EBV, and could guide the development of novel MS therapies.

  View details for DOI 10.1038/s41586-022-04432-7

  View details for PubMedID 35073561

• Epstein-Barr virus and multiple sclerosis. Science (New York, N.Y.) Robinson, W. H., Steinman, L. 2022; 375 (6578): 264-265

  Abstract

  [Figure: see text].

  View details for DOI 10.1126/science.abm7930

  View details for PubMedID 35025606

• BNT162b2 vaccine induces divergent B cell responses to SARS-CoV-2 S1 and S2 NATURE IMMUNOLOGY Brewer, R., Ramadoss, N. S., Lahey, L. J., Jahanbani, S., Robinson, W. H., Lanz, T. 2021

  Abstract

  The first ever US Food and Drug Administration-approved messenger RNA vaccines are highly protective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)1-3. However, the contribution of each dose to the generation of antibodies against SARS-CoV-2 spike (S) protein and the degree of protection against novel variants warrant further study. Here, we investigated the B cell response to the BNT162b2 vaccine by integrating B cell repertoire analysis with single-cell transcriptomics pre- and post-vaccination. The first vaccine dose elicits a recall response of IgA+ plasmablasts targeting the S subunit S2. Three weeks after the first dose, we observed an influx of minimally mutated IgG+ memory B cells that targeted the receptor binding domain on the S subunit S1 and likely developed from the naive B cell pool. This response was strongly boosted by the second dose and delivers potently neutralizing antibodies against SARS-CoV-2 and several of its variants.

  View details for DOI 10.1038/s41590-021-01088-9

  View details for Web of Science ID 000723985200001

  View details for PubMedID 34848871

• Neutralizing Anti-Interleukin-1 Receptor-Antagonist Autoantibodies Induce Inflammatory and Fibrotic Mediators in IgG4-Related Disease. The Journal of allergy and clinical immunology Jarrell, J. A., Baker, M. C., Perugino, C. A., Liu, H., Bloom, M. S., Maehara, T., Wong, H. H., Lanz, T., Adamska, J. Z., Kongpachith, S., Sokolove, J., Stone, J. H., Pillai, S. S., Robinson, W. H. 2021

  Abstract

  BACKGROUND: IgG4-related disease (IgG4-RD) is a fibro-inflammatory condition involving loss of B cell tolerance and production of autoantibodies. However, the relevant targets and role of these aberrant humoral immune responses are not defined.OBJECTIVE: To identify novel autoantibodies and autoantigen targets that promote pathogenic responses in IgG4-RD.METHODS: We sequenced plasmablast antibody repertoires in patients with IgG4-RD. Representative monoclonal antibodies (mAb) were expressed and their specificities characterized using cytokine microarrays. The role of anti-interleukin-1 receptor-antagonist (IL-1RA) autoantibodies was investigated using in vitro assays.RESULTS: We identified strong reactivity against human IL-1RA using a clonally-expanded plasmablast-derived mAb from a patient with IgG4-RD. IgG4-RD patient plasma exhibited elevated levels of reactivity against IL-1RA compared to controls and neutralized IL-1RA activity, resulting in inflammatory and fibrotic mediator production in vitro. IL-1RA was detected in lesional tissues from IgG4-RD patients. Patients with anti-IL-1RA autoantibodies of the IgG4 subclass had greater numbers of organs affected than those without anti-IL-1RA autoantibodies. Peptide analyses identified IL-1RA epitopes targeted by anti-IL-1RA antibodies at sites near the IL-1RA/IL-1R interface. Serum from patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) also had elevated levels of anti-IL-1RA autoantibodies compared to controls.CONCLUSION: A subset of patients with IgG4-RD have anti-IL-1RA autoantibodies, which promote pro-inflammatory and pro-fibrotic meditator production via IL-1RA neutralization. These findings support a novel immunological mechanism underlying the pathogenesis of IgG4-RD. Anti-IL-1RA autoantibodies are also present in a subset of patients with SLE and RA, suggesting a potential common pathway in multiple autoimmune diseases.

  View details for DOI 10.1016/j.jaci.2021.05.002

  View details for PubMedID 33974929

• IgE-mediated mast cell activation promotes inflammation and cartilage destruction in osteoarthritis ELIFE Wang, Q., Lepus, C. M., Raghu, H., Reber, L. L., Tsai, M. M., Wong, H. H., von Kaeppler, E., Lingampalli, N., Bloom, M. S., Hu, N., Elliott, E. E., Oliviero, F., Punzi, L., Giori, N. J., Goodman, S. B., Chu, C. R., Sokolove, J., Fukuoka, Y., Schwartz, L. B., Galli, S. J., Robinson, W. H. 2019; 8

  View details for DOI 10.7554/eLife.39905

  View details for Web of Science ID 000467895900001

• Affinity Maturation of the Anti-Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Kongpachith, S., Lingampalli, N., Ju, C., Blum, L. K., Lu, D. R., Elliott, S. E., Mao, R., Robinson, W. H. 2019; 71 (4): 507–17

  View details for DOI 10.1002/art.40760

  View details for Web of Science ID 000464454700004

• IgE-mediated mast cell activation promotes inflammation and cartilage destruction in osteoarthritis. eLife Wang, Q. n., Lepus, C. M., Raghu, H. n., Reber, L. L., Tsai, M. M., Wong, H. H., von Kaeppler, E. n., Lingampalli, N. n., Bloom, M. S., Hu, N. n., Elliott, E. E., Oliviero, F. n., Punzi, L. n., Giori, N. J., Goodman, S. B., Chu, C. R., Sokolove, J. n., Fukuoka, Y. n., Schwartz, L. B., Galli, S. J., Robinson, W. H. 2019; 8

  Abstract

  Osteoarthritis is characterized by articular cartilage breakdown, and emerging evidence suggests that dysregulated innate immunity is likely involved. Here, we performed proteomic, transcriptomic, and electron microscopic analyses to demonstrate that mast cells are aberrantly activated in human and murine osteoarthritic joint tissues. Using genetic models of mast cell deficiency, we demonstrate that lack of mast cells attenuates osteoarthritis in mice. Using genetic and pharmacologic approaches, we show that the IgE/FcεRI/Syk signaling axis is critical for the development of osteoarthritis. We find that mast cell-derived tryptase induces inflammation, chondrocyte apoptosis, and cartilage breakdown. Our findings demonstrate a central role for IgE-dependent mast cell activation in the pathogenesis of osteoarthritis, suggesting that targeting mast cells could provide therapeutic benefit in human osteoarthritis.This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review. The Reviewing Editor's assessment is that all the issues have been addressed (see decision letter).

  View details for PubMedID 31084709

• Dysregulated integrin αVβ3 and CD47 signaling promotes joint inflammation, cartilage breakdown, and progression of osteoarthritis. JCI insight Wang, Q. n., Onuma, K. n., Liu, C. n., Wong, H. n., Bloom, M. S., Elliott, E. E., Cao, R. R., Hu, N. n., Lingampalli, N. n., Sharpe, O. n., Zhao, X. n., Sohn, D. H., Lepus, C. M., Sokolove, J. n., Mao, R. n., Cisar, C. T., Raghu, H. n., Chu, C. R., Giori, N. J., Willingham, S. B., Prohaska, S. S., Cheng, Z. n., Weissman, I. L., Robinson, W. H. 2019; 4 (18)

  Abstract

  Osteoarthritis (OA) is the leading cause of joint failure, yet the underlying mechanisms remain elusive, and no approved therapies that slow progression exist. Dysregulated integrin function was previously implicated in OA pathogenesis. However, the roles of integrin αVβ3 and the integrin-associated receptor CD47 in OA remain largely unknown. Here, transcriptomic and proteomic analyses of human and murine osteoarthritic tissues revealed dysregulated expression of αVβ3, CD47, and their ligands. Using genetically deficient mice and pharmacologic inhibitors, we showed that αVβ3, CD47, and the downstream signaling molecules Fyn and FAK are crucial to OA pathogenesis. MicroPET/CT imaging of a mouse model showed elevated ligand-binding capacities of integrin αVβ3 and CD47 in osteoarthritic joints. Further, our in vitro studies demonstrated that chondrocyte breakdown products, derived from articular cartilage of individuals with OA, induced αVβ3/CD47-dependent expression of inflammatory and degradative mediators, and revealed the downstream signaling network. Our findings identify a central role for dysregulated αVβ3 and CD47 signaling in OA pathogenesis and suggest that activation of αVβ3 and CD47 signaling in many articular cell types contributes to inflammation and joint destruction in OA. Thus, the data presented here provide a rationale for targeting αVβ3, CD47, and their signaling pathways as a disease-modifying therapy.

  View details for DOI 10.1172/jci.insight.128616

  View details for PubMedID 31534047

• Affinity Maturation Drives Epitope Spreading and Generation of Pro-inflammatory Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis. Arthritis & rheumatology (Hoboken, N.J.) Elliott, S. E., Kongpachith, S., Lingampalli, N., Adamska, J. Z., Cannon, B. J., Mao, R., Blum, L. K., Robinson, W. H. 2018

  Abstract

  OBJECTIVE: Rheumatoid arthritis (RA) is characterized by anti-citrullinated protein antibodies (ACPAs), nevertheless, the origin, specificity, and functional properties of ACPAs remain poorly understood. To characterize the evolution of ACPAs, we sequenced the plasmablast antibody repertoire at serial timepoints in subjects with established RA.METHODS: Blood samples were obtained at up to four timepoints from eight anti-CCP+ individuals with established RA. We single-cell sorted CD19+CD3-IgD-CD14-CD20-CD27+CD38++ plasmablasts and co-stained with citrullinated-peptide tetramers to identify ACPA-expressing plasmablasts. Cell-specific oligonucleotide barcodes were utilized followed by large-scale sequencing and bioinformatic analysis to obtain error-corrected, paired, heavy and light chain antibody gene sequences for each B cell.RESULTS: Bioinformatic analysis revealed 170 persistent plasmablast lineages of which 19% included multiple isotypes. Among IgG- and IgA-expressing plasmablasts, we observed significantly more IgA-expressing persistent lineages compared to IgG (P < 0.01). We identified shared CDR3 sequence motifs across subjects. A subset of lineages comprised of later-timepoint derived members with divergent somatic hypermutations encoded antibodies that bind an expanded set of citrullinated antigens. Further, these recombinant, differentially mutated plasmablast antibodies formed immune complexes that stimulated higher levels of macrophage TNF-alpha production compared to antibodies representing earlier-timepoint, less-mutated lineage members.CONCLUSIONS: Our findings demonstrate that established RA is characterized by a persistent IgA ACPA response that exhibits ongoing affinity maturation. This observation suggests the presence of a persistent mucosal antigen that continually promotes the production of IgA plasmablasts, and their affinity maturation and epitope spreading to generate ACPAs that bind additional citrullinated antigens and more potently stimulate macrophage TNF-alpha production. This article is protected by copyright. All rights reserved.

  View details for PubMedID 29927104

• T cell-dependent affinity maturation and innate immune pathways differentially drive autoreactive B cell responses in rheumatoid arthritis. Arthritis & rheumatology (Hoboken, N.J.) Lu, D. R., McDavid, A. N., Kongpachith, S., Lingampalli, N., Glanville, J., Ju, C., Gottardo, R., Robinson, W. H. 2018

  Abstract

  OBJECTIVE: Rheumatoid arthritis (RA) is characterized by the activation of B cells that produce anti-citrullinated protein antibodies (ACPA) and rheumatoid factors (RF), but the mechanisms by which tolerance is broken in these B cells remain incompletely understood. Here we investigate whether ACPA and RF B cells break tolerance through distinct molecular mechanisms.METHOD: We developed antigen-tetramers to isolate ACPA- and RF-producing B cells and performed single-cell RNA-sequencing on 2,349 B cells from six RA patients to analyze their immunoglobulin repertoires and transcriptional programs. Prominent immunoglobulins were expressed as monoclonal antibodies and tested for autoantigen reactivity.RESULTS: ACPA and RF B cells were enriched in the peripheral blood of RA patients relative to healthy controls. Characterization of patient-derived monoclonal antibodies confirmed ACPA and RF targeting of tetramer-specific B cells at both antigen-inexperienced and affinity-matured B-cell stages. ACPA B cells utilized more class-switched isotypes and exhibited more somatic hypermutations relative to RF B cells, and these differences were accompanied by the downregulation of CD72 and upregulation of genes that promote class-switching and T cell-dependent responses. In contrast, RF B cells expressed transcriptional programs that stimulate rapid memory reactivation through multiple innate immune pathways. Coexpression analysis revealed that ACPA- and RF- B cell enriched genes belong to distinct transcriptional regulatory networks.CONCLUSION: Our findings suggest that ACPA and RF B cells are imprinted with distinct transcriptional programs, suggesting that these autoantibodies associated with increased inflammation in RA arise from two different molecular mechanisms. This article is protected by copyright. All rights reserved.

  View details for PubMedID 29855173

• Non-progressing cancer patients have persistent B cell responses expressing shared antibody paratopes that target public tumor antigens CLINICAL IMMUNOLOGY DeFalco, J., Harbell, M., Manning-Bog, A., Baia, G., Scholz, A., Millare, B., Sumi, M., Zhang, D., Chu, F., Dowd, C., Zuno-Mitchell, P., Kim, D., Leung, Y., Jiang, S., Tang, X., Williamson, K. S., Chen, X., Carroll, S. M., Santo, G., Haaser, N., Ngan Nguyen, Giladi, E., Minor, D., Tan, Y., Sokolove, J. B., Steinman, L., Serafini, T. A., Cavet, G., Greenberg, N. M., Glanville, J., Volkmuth, W., Emerling, D. E., Robinson, W. H. 2018; 187: 37–45

  Abstract

  There is significant debate regarding whether B cells and their antibodies contribute to effective anti-cancer immune responses. Here we show that patients with metastatic but non-progressing melanoma, lung adenocarcinoma, or renal cell carcinoma exhibited increased levels of blood plasmablasts. We used a cell-barcoding technology to sequence their plasmablast antibody repertoires, revealing clonal families of affinity matured B cells that exhibit progressive class switching and persistence over time. Anti-CTLA4 and other treatments were associated with further increases in somatic hypermutation and clonal family size. Recombinant antibodies from clonal families bound non-autologous tumor tissue and cell lines, and families possessing immunoglobulin paratope sequence motifs shared across patients exhibited increased rates of binding. We identified antibodies that caused regression of, and durable immunity toward, heterologous syngeneic tumors in mice. Our findings demonstrate convergent functional anti-tumor antibody responses targeting public tumor antigens, and provide an approach to identify antibodies with diagnostic or therapeutic utility.

  View details for PubMedID 29031828

• Barcode-Enabled Sequencing of Plasmablast Antibody Repertoires in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Tan, Y., Kongpachith, S., Blum, L. K., Ju, C., Lahey, L. J., Lu, D. R., Cai, X., Wagner, C. A., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 66 (10): 2706-2715

  Abstract

  A hallmark of rheumatoid arthritis (RA) is the production of autoantibodies, including anti-citrullinated protein antibodies (ACPAs). Nevertheless, the specific targets of these autoantibodies remain incompletely defined. During an immune response, B cells specific for the inciting antigen(s) are activated and differentiate into plasmablasts, which are released into the blood. We undertook this study to sequence the plasmablast antibody repertoire to define the targets of the active immune response in RA.We developed a novel DNA barcoding method to sequence the cognate heavy- and light-chain pairs of antibodies expressed by individual blood plasmablasts in RA. The method uses a universal 5' adapter that enables full-length sequencing of the antibodies' variable regions and recombinant expression of the paired antibody chains. The sequence data sets were bioinformatically analyzed to generate phylogenetic trees that identify clonal families of antibodies sharing heavy- and light-chain VJ sequences. Representative antibodies were expressed, and their binding properties were characterized using anti-cyclic citrullinated peptide 2 (anti-CCP-2) enzyme-linked immunosorbent assay (ELISA) and antigen microarrays.We used our sequencing method to generate phylogenetic trees representing the antibody repertoires of peripheral blood plasmablasts from 4 individuals with anti-CCP+ RA, and recombinantly expressed 14 antibodies that were either "singleton" antibodies or representative of clonal antibody families. Anti-CCP-2 ELISA identified 4 ACPAs, and antigen microarray analysis identified ACPAs that differentially targeted epitopes on α-enolase, citrullinated fibrinogen, and citrullinated histone H2B.Our data provide evidence that autoantibodies targeting α-enolase, citrullinated fibrinogen, and citrullinated histone H2B are produced by the ongoing activated B cell response in, and thus may contribute to the pathogenesis of, RA.

  View details for DOI 10.1002/art.38754

  View details for Web of Science ID 000342744300008

  View details for PubMedCentralID PMC4560105

• High-throughput sequencing of natively paired antibody chains provides evidence for original antigenic sin shaping the antibody response to influenza vaccination. Clinical immunology Tan, Y., Blum, L. K., Kongpachith, S., Ju, C., Cai, X., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 151 (1): 55-65

  Abstract

  We developed a DNA barcoding method to enable high-throughput sequencing of the cognate heavy- and light-chain pairs of the antibodies expressed by individual B cells. We used this approach to elucidate the plasmablast antibody response to influenza vaccination. We show that >75% of the rationally selected plasmablast antibodies bind and neutralize influenza, and that antibodies from clonal families, defined by sharing both heavy-chain VJ and light-chain VJ sequence usage, do so most effectively. Vaccine-induced heavy-chain VJ regions contained on average >20 nucleotide mutations as compared to their predicted germline gene sequences, and some vaccine-induced antibodies exhibited higher binding affinities for hemagglutinins derived from prior years' seasonal influenza as compared to their affinities for the immunization strains. Our results show that influenza vaccination induces the recall of memory B cells that express antibodies that previously underwent affinity maturation against prior years' seasonal influenza, suggesting that 'original antigenic sin' shapes the antibody response to influenza vaccination.

  View details for DOI 10.1016/j.clim.2013.12.008

  View details for PubMedID 24525048

• Identification of a central role for complement in osteoarthritis NATURE MEDICINE Wang, Q., Rozelle, A. L., Lepus, C. M., Scanzello, C. R., Song, J. J., Larsen, D. M., Crish, J. F., Bebek, G., Ritter, S. Y., Lindstrom, T. M., Hwang, I., Wong, H. H., Punzi, L., Encarnacion, A., Shamloo, M., Goodman, S. B., Wyss-Coray, T., Goldring, S. R., Banda, N. K., Thurman, J. M., Gobezie, R., Crow, M. K., Holers, V. M., Lee, D. M., Robinson, W. H. 2011; 17 (12): 1674-U196

  Abstract

  Osteoarthritis, characterized by the breakdown of articular cartilage in synovial joints, has long been viewed as the result of 'wear and tear'. Although low-grade inflammation is detected in osteoarthritis, its role is unclear. Here we identify a central role for the inflammatory complement system in the pathogenesis of osteoarthritis. Through proteomic and transcriptomic analyses of synovial fluids and membranes from individuals with osteoarthritis, we find that expression and activation of complement is abnormally high in human osteoarthritic joints. Using mice genetically deficient in complement component 5 (C5), C6 or the complement regulatory protein CD59a, we show that complement, specifically, the membrane attack complex (MAC)-mediated arm of complement, is crucial to the development of arthritis in three different mouse models of osteoarthritis. Pharmacological modulation of complement in wild-type mice confirmed the results obtained with genetically deficient mice. Expression of inflammatory and degradative molecules was lower in chondrocytes from destabilized joints from C5-deficient mice than C5-sufficient mice, and MAC induced production of these molecules in cultured chondrocytes. Further, MAC colocalized with matrix metalloprotease 13 (MMP13) and with activated extracellular signal-regulated kinase (ERK) around chondrocytes in human osteoarthritic cartilage. Our findings indicate that dysregulation of complement in synovial joints has a key role in the pathogenesis of osteoarthritis.

  View details for DOI 10.1038/nm.2543

  View details for Web of Science ID 000297978000042

  View details for PubMedID 22057346

  View details for PubMedCentralID PMC3257059

• Selective tyrosine kinase inhibition by imatinib mesylate for the treatment of autoimmune arthritis JOURNAL OF CLINICAL INVESTIGATION Paniagua, R. T., Sharpe, O., Ho, P. P., Chan, S. M., Chang, A., Higgins, J. P., Tomooka, B. H., Thomas, F. M., Song, J. J., Goodman, S. B., Lee, D. M., Genovese, M. C., Utz, P. J., Steinman, L., Robinson, W. H. 2006; 116 (10): 2633-2642

  Abstract

  Tyrosine kinases play a central role in the activation of signal transduction pathways and cellular responses that mediate the pathogenesis of rheumatoid arthritis. Imatinib mesylate (imatinib) is a tyrosine kinase inhibitor developed to treat Bcr/Abl-expressing leukemias and subsequently found to treat c-Kit-expressing gastrointestinal stromal tumors. We demonstrate that imatinib potently prevents and treats murine collagen-induced arthritis (CIA). We further show that micromolar concentrations of imatinib abrogate multiple signal transduction pathways implicated in RA pathogenesis, including mast cell c-Kit signaling and TNF-alpha release, macrophage c-Fms activation and cytokine production, and fibroblast PDGFR signaling and proliferation. In our studies, imatinib attenuated PDGFR signaling in fibroblast-like synoviocytes (FLSs) and TNF-alpha production in synovial fluid mononuclear cells (SFMCs) derived from human RA patients. Imatinib-mediated inhibition of a spectrum of signal transduction pathways and the downstream pathogenic cellular responses may provide a powerful approach to treat RA and other inflammatory diseases.

  View details for DOI 10.1172/JCI28546

  View details for PubMedID 16981009

• Lipid microarrays identify key mediators of autoimmune brain inflammation NATURE MEDICINE Kanter, J. L., Narayana, S., Ho, P. P., Catz, I., Warren, K. G., Sobel, R. A., Steinman, L., Robinson, W. H. 2006; 12 (1): 138-143

  Abstract

  Recent studies suggest that increased T-cell and autoantibody reactivity to lipids may be present in the autoimmune demyelinating disease multiple sclerosis. To perform large-scale multiplex analysis of antibody responses to lipids in multiple sclerosis, we developed microarrays composed of lipids present in the myelin sheath, including ganglioside, sulfatide, cerebroside, sphingomyelin and total brain lipid fractions. Lipid-array analysis showed lipid-specific antibodies against sulfatide, sphingomyelin and oxidized lipids in cerebrospinal fluid (CSF) derived from individuals with multiple sclerosis. Sulfatide-specific antibodies were also detected in SJL/J mice with acute experimental autoimmune encephalomyelitis (EAE). Immunization of mice with sulfatide plus myelin peptide resulted in a more severe disease course of EAE, and administration of sulfatide-specific antibody exacerbated EAE. Thus, autoimmune responses to sulfatide and other lipids are present in individuals with multiple sclerosis and in EAE, and may contribute to the pathogenesis of autoimmune demyelination.

  View details for DOI 10.1038/nm1344

  View details for PubMedID 16341241

• Protein microarrays guide tolerizing DNA vaccine treatment of autoimmune encephalomyelitis NATURE BIOTECHNOLOGY Robinson, W. H., Fontoura, P., Lee, B. J., de Vegvar, H. E., Tom, J., Pedotti, R., DiGennaro, C. D., Mitchell, D. J., Fong, D., Ho, P. P., Ruiz, P. J., Maverakis, E., Stevens, D. B., Bernard, C. C., Martin, R., Kuchroo, V. K., van Noort, J. M., Genain, C. P., Amor, S., Olsson, T., Utz, P. J., Garren, H., Steinman, L. 2003; 21 (9): 1033-1039

  Abstract

  The diversity of autoimmune responses poses a formidable challenge to the development of antigen-specific tolerizing therapy. We developed 'myelin proteome' microarrays to profile the evolution of autoantibody responses in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis (MS). Increased diversity of autoantibody responses in acute EAE predicted a more severe clinical course. Chronic EAE was associated with previously undescribed extensive intra- and intermolecular epitope spreading of autoreactive B-cell responses. Array analysis of autoantigens targeted in acute EAE was used to guide the choice of autoantigen cDNAs to be incorporated into expression plasmids so as to generate tolerizing vaccines. Tolerizing DNA vaccines encoding a greater number of array-determined myelin targets proved superior in treating established EAE and reduced epitope spreading of autoreactive B-cell responses. Proteomic monitoring of autoantibody responses provides a useful approach to monitor autoimmune disease and to develop and tailor disease- and patient-specific tolerizing DNA vaccines.

  View details for DOI 10.1038/nbt859

  View details for PubMedID 12910246

• Autoantigen microarrays for multiplex characterization of autoantibody responses NATURE MEDICINE Robinson, W. H., DiGennaro, C., Hueber, W., Haab, B. B., KAMACHI, M., Dean, E. J., Fournel, S., Fong, D., Genovese, M. C., de Vegvar, H. E., Skriner, K., Hirschberg, D. L., Morris, R. I., Muller, S., Pruijn, G. J., van Venrooij, W. J., Smolen, J. S., Brown, P. O., Steinman, L., Utz, P. J. 2002; 8 (3): 295-301

  Abstract

  We constructed miniaturized autoantigen arrays to perform large-scale multiplex characterization of autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases.

  View details for Web of Science ID 000174139500036

  View details for PubMedID 11875502

• Dysregulated fibrinolysis and plasmin activation promote the pathogenesis of osteoarthritis. JCI insight Wang, Q., Shao, G., Zhao, X., Wong, H. H., Chin, K., Zhao, M., Bai, A., Bloom, M. S., Love, Z. Z., Chu, C. R., Cheng, Z., Robinson, W. H. 2024

  Abstract

  Joint injury is associated with risk for development of osteoarthritis (OA). Increasing evidence suggests that activation of fibrinolysis is involved in OA pathogenesis. However, the role of the fibrinolytic pathway is not well understood. Here we showed that the fibrinolytic pathway, which includes plasminogen/plasmin, tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA) and the uPA receptor (uPAR), were dysregulated in human OA joints. Pharmacological inhibition of plasmin attenuated OA progression in a destabilization of the medial meniscus (DMM) mouse model, while genetic deficiency of plasmin activator inhibitor (PAI-1), or injection of plasmin, exacerbated OA. We detected increased uptake of uPA/uPAR in mouse OA joints by microPET/CT imaging. In vitro studies identified that plasmin promotes OA development through multiple mechanisms, including the degradation of lubricin and cartilage proteoglycans, induction of inflammatory and degradative mediators. We showed that uPA and uPAR produced inflammatory and degradative mediators by activating the PI3K, PDK1, AKT, and ERK signaling cascades, and activates matrix metalloproteinases (pro-MMPs) to degrade proteoglycan. Together, we demonstrated that fibrinolysis contributes to the development of OA through multiple mechanisms and suggested that therapeutic targeting of the fibrinolysis pathway can prevent or slow development of OA.

  View details for DOI 10.1172/jci.insight.173603

  View details for PubMedID 38502232

• Expansion of HLA-DR+ Peripheral Helper T and Naïve B cells in Anti-Citrullinated Protein Antibody-Positive Subjects At-Risk for Rheumatoid Arthritis. Arthritis & rheumatology (Hoboken, N.J.) Takada, H., Demoruelle, M. K., Deane, K. D., Nakamura, S., Katsumata, Y., Ikari, K., Buckner, J. H., Robinson, W. H., Seifert, J. A., Feser, M. L., Moss, L., Norris, J. M., Harigai, M., Hsieh, E. W., Holers, V. M., Okamoto, Y. 2024

  Abstract

  To investigate immune dysregulation in the peripheral blood that contributes to the pre-rheumatoid arthritis (RA) stage of RA development in anti-citrullinated protein antibody (ACPA)+ individuals.Using 37 markers by mass cytometry, we investigated peripheral blood mononuclear cells (PBMCs) from ACPA+ at-risk individuals (ARI), ACPA+ early untreated RA patients, and ACPA- controls in the Tokyo Women's Medical University (TWMU) cohort (n = 17 in each group). Computational algorithms, FlowSOM and Optimized t-Distributed Stochastic Neighbor Embedding (opt-SNE), were employed to explore specific immunological differences between study groups. These findings were further evaluated, and longitudinal changes were explored, using flow cytometry and PBMCs from the USA-based Targeting Immune Responses for Prevention of RA (TIP-RA) cohort that included 11 ACPA+ individuals who later developed RA (pre-RA), of which 9 had post-RA diagnosis PBMCs (post-RA), and 11 ACPA- controls.HLA-DR+ Tph cells, activated regulatory T cells, PD-1hi CD8+ T cells, and CXCR5- CD11c- CD38+ naïve B cells were significantly expanded in PBMCs from ARI and early RA patients from the TWMU cohort. Expansion of HLA-DR+ Tph cells and CXCR5- CD11c- CD38+ naïve B cells was likewise found in both pre-RA and post-RA time points in the TIP-RA cohort.The expansion of HLA-DR+ Tph cells and CXCR5- CD11c- CD38+ naïve B cells in ACPA+ individuals, including those who developed inflammatory arthritis and classified RA, supports a key role of these cells in transition from pre-RA to classified RA. These findings may identify a new mechanistic target for treatment and prevention in RA.

  View details for DOI 10.1002/art.42839

  View details for PubMedID 38412870

• Longitudinal patterns and predictors of response to standard-of-care therapy in lupus nephritis: data from the Accelerating Medicines Partnership Lupus Network. Arthritis research & therapy Izmirly, P. M., Kim, M. Y., Carlucci, P. M., Preisinger, K., Cohen, B. Z., Deonaraine, K., Zaminski, D., Dall'Era, M., Kalunian, K., Fava, A., Belmont, H. M., Wu, M., Putterman, C., Anolik, J., Barnas, J. L., Diamond, B., Davidson, A., Wofsy, D., Kamen, D., James, J. A., Guthridge, J. M., Apruzzese, W., Rao, D. A., Weisman, M. H., Petri, M., Buyon, J., Furie, R. 2024; 26 (1): 54

  Abstract

  Leveraging the Accelerating Medicines Partnership (AMP) Lupus Nephritis (LN) dataset, we evaluated longitudinal patterns, rates, and predictors of response to standard-of-care therapy in patients with lupus nephritis.Patients from US academic medical centers with class III, IV, and/or V LN and a baseline urine protein/creatinine (UPCR) ratio ≥ 1.0 (n = 180) were eligible for this analysis. Complete response (CR) required the following: (1) UPCR < 0.5; (2) normal serum creatinine (≤ 1.3 mg/dL) or, if abnormal, ≤ 125% of baseline; and (3) prednisone ≤ 10 mg/day. Partial response (PR) required the following: (1) > 50% reduction in UPCR; (2) normal serum creatinine or, if abnormal, ≤ 125% of baseline; and (3) prednisone dose ≤ 15 mg/day.Response rates to the standard of care at week 52 were CR = 22.2%; PR = 21.7%; non-responder (NR) = 41.7%, and not determined (ND) = 14.4%. Only 8/180 (4.4%) patients had a week 12 CR sustained through week 52. Eighteen (10%) patients attained a week 12 PR or CR and sustained their responses through week 52 and 47 (26.1%) patients achieved sustained PR or CR at weeks 26 and 52. Week 52 CR or PR attainment was associated with baseline UPCR > 3 (ORadj = 3.71 [95%CI = 1.34-10.24]; p = 0.012), > 25% decrease in UPCR from baseline to week 12 (ORadj = 2.61 [95%CI = 1.07-6.41]; p = 0.036), lower chronicity index (ORadj = 1.33 per unit decrease [95%CI = 1.10-1.62]; p = 0.003), and positive anti-dsDNA antibody (ORadj = 2.61 [95%CI = 0.93-7.33]; p = 0.069).CR and PR rates at week 52 were consistent with the standard-of-care response rates observed in prospective registrational LN trials. Low sustained response rates underscore the need for more efficacious therapies and highlight how critically important it is to understand the molecular pathways associated with response and non-response.

  View details for DOI 10.1186/s13075-024-03275-z

  View details for PubMedID 38378664

  View details for PubMedCentralID 3409311

• Immune tolerance of citrullinated peptides. Nature reviews. Rheumatology Thomas, R., Robinson, W. H. 2024

  View details for DOI 10.1038/s41584-024-01081-0

  View details for PubMedID 38263304

  View details for PubMedCentralID 3832918

• A candidate antibody drug for prevention of malaria. Nature medicine Williams, K. L., Guerrero, S., Flores-Garcia, Y., Kim, D., Williamson, K. S., Siska, C., Smidt, P., Jepson, S. Z., Li, K., Dennison, S. M., Mathis-Torres, S., Chen, X., Wille-Reece, U., MacGill, R. S., Walker, M., Jongert, E., King, C. R., Ockenhouse, C., Glanville, J., Moon, J. E., Regules, J. A., Tan, Y. C., Cavet, G., Lippow, S. M., Robinson, W. H., Dutta, S., Tomaras, G. D., Zavala, F., Ketchem, R. R., Emerling, D. E. 2024

  Abstract

  Over 75% of malaria-attributable deaths occur in children under the age of 5 years. However, the first malaria vaccine recommended by the World Health Organization (WHO) for pediatric use, RTS,S/AS01 (Mosquirix), has modest efficacy. Complementary strategies, including monoclonal antibodies, will be important in efforts to eradicate malaria. Here we characterize the circulating B cell repertoires of 45 RTS,S/AS01 vaccinees and discover monoclonal antibodies for development as potential therapeutics. We generated >28,000 antibody sequences and tested 481 antibodies for binding activity and 125 antibodies for antimalaria activity in vivo. Through these analyses we identified correlations suggesting that sequences in Plasmodium falciparum circumsporozoite protein, the target antigen in RTS,S/AS01, may induce immunodominant antibody responses that limit more protective, but subdominant, responses. Using binding studies, mouse malaria models, biomanufacturing assessments and protein stability assays, we selected AB-000224 and AB-007088 for advancement as a clinical lead and backup. We engineered the variable domains (Fv) of both antibodies to enable low-cost manufacturing at scale for distribution to pediatric populations, in alignment with WHO's preferred product guidelines. The engineered clone with the optimal manufacturing and drug property profile, MAM01, was advanced into clinical development.

  View details for DOI 10.1038/s41591-023-02659-z

  View details for PubMedID 38167935

  View details for PubMedCentralID 5395940

• Multifaceted immune dysregulation characterizes individuals at-risk for rheumatoid arthritis. Nature communications James, E. A., Holers, V. M., Iyer, R., Prideaux, E. B., Rao, N. L., Rims, C., Muir, V. S., Posso, S. E., Bloom, M. S., Zia, A., Elliott, S. E., Adamska, J. Z., Ai, R., Brewer, R. C., Seifert, J. A., Moss, L., Barzideh, S., Demoruelle, M. K., Striebich, C. C., Okamoto, Y., Sainbayar, E., Crook, A. A., Peterson, R. A., Vanderlinden, L. A., Wang, W., Boyle, D. L., Robinson, W. H., Buckner, J. H., Firestein, G. S., Deane, K. D. 2023; 14 (1): 7637

  Abstract

  Molecular markers of autoimmunity, such as antibodies to citrullinated protein antigens (ACPA), are detectable prior to inflammatory arthritis (IA) in rheumatoid arthritis (RA) and may define a state that is 'at-risk' for future RA. Here we present a cross-sectional comparative analysis among three groups that include ACPA positive individuals without IA (At-Risk), ACPA negative individuals and individuals with early, ACPA positive clinical RA (Early RA). Differential methylation analysis among the groups identifies non-specific dysregulation in peripheral B, memory and naive T cells in At-Risk participants, with more specific immunological pathway abnormalities in Early RA. Tetramer studies show increased abundance of T cells recognizing citrullinated (cit) epitopes in At-Risk participants, including expansion of T cells reactive to citrullinated cartilage intermediate layer protein I (cit-CILP); these T cells have Th1, Th17, and T stem cell memory-like phenotypes. Antibody-antigen array analyses show that antibodies targeting cit-clusterin, cit-fibrinogen and cit-histone H4 are elevated in At-Risk and Early RA participants, with the highest levels of antibodies detected in those with Early RA. These findings indicate that an ACPA positive at-risk state is associated with multifaceted immune dysregulation that may represent a potential opportunity for targeted intervention.

  View details for DOI 10.1038/s41467-023-43091-8

  View details for PubMedID 37993439

• A novel mouse model of cerebral adrenoleukodystrophy highlights NLRP3 activity in lesion pathogenesis. bioRxiv : the preprint server for biology Hashemi, E., Narain Srivastava, I., Aguirre, A., Tilahan Yoseph, E., Kaushal, E., Awani, A., Kyu Ryu, J., Akassoglou, K., Talebian, S., Chu, P., Pisani, L., Musolino, P., Steinman, L., Doyle, K., Robinson, W. H., Sharpe, O., Cayrol, R., Orchard, P., Lund, T., Vogel, H., Lenail, M., Han, M. H., Bonkowsky, J. L., Van Haren, K. P. 2023

  Abstract

  We sought to create and characterize a mouse model of the inflammatory, cerebral demyelinating phenotype of X-linked adrenoleukodystrophy (ALD) that would facilitate the study of disease pathogenesis and therapy development. We also sought to cross-validate potential therapeutic targets such as fibrin, oxidative stress, and the NLRP3 inflammasome, in post-mortem human and murine brain tissues.ALD is caused by mutations in the gene ABCD1 encoding a peroxisomal transporter. More than half of males with an ABCD1 mutation develop the cerebral phenotype (cALD). Incomplete penetrance and absence of a genotype-phenotype correlation imply a role for environmental triggers. Mechanistic studies have been limited by the absence of a cALD phenotype in the Abcd1-null mouse.We generated a cALD phenotype in 8-week-old, male Abcd1-null mice by deploying a two-hit method that combines cuprizone (CPZ) and experimental autoimmune encephalomyelitis (EAE) models. We employed in vivo MRI and post-mortem immunohistochemistry to evaluate myelin loss, astrogliosis, blood-brain barrier (BBB) disruption, immune cell infiltration, fibrin deposition, oxidative stress, and Nlrp3 inflammasome activation in mice. We used bead-based immunoassay and immunohistochemistry to evaluate IL-18 in CSF and post-mortem human cALD brain tissue.MRI studies revealed T2 hyperintensities and post-gadolinium enhancement in the medial corpus callosum of cALD mice, similar to human cALD lesions. Both human and mouse cALD lesions shared common histologic features of myelin phagocytosis, myelin loss, abundant microglial activation, T and B-cell infiltration, and astrogliosis. Compared to wild-type controls, Abcd1-null mice had more severe cerebral inflammation, demyelination, fibrin deposition, oxidative stress, and IL-18 activation. IL-18 immunoreactivity co-localized with macrophages/microglia in the perivascular region of both human and mouse brain tissue.This novel mouse model of cALD suggests loss of Abcd1 function predisposes to more severe cerebral inflammation, oxidative stress, fibrin deposition, and Nlrp3 pathway activation, which parallels the findings seen in humans with cALD. We expect this model to enable long-sought investigations into cALD mechanisms and accelerate development of candidate therapies for lesion prevention, cessation, and remyelination.

  View details for DOI 10.1101/2023.11.07.564025

  View details for PubMedID 37986739

  View details for PubMedCentralID PMC10659266

• Genetic association between atopic disease and osteoarthritis. Osteoarthritis and cartilage Baker, M. C., Robinson, W. H., Ostrom, Q. 2023

  Abstract

  To evaluate the association between genetically determined risk for atopic disease and osteoarthritis (OA).We performed linkage disequilibrium (LD) score regression (LDSC) using 1,000 Genomes Project European samples as a reference for patterns of genome-wide LD. Summary statistics for atopic disease traits were obtained from the UK Biobank. We generated pairwise genetic correlation between OA and traits for atopic disease to estimate genetic correlation between traits (rg) and heritability for each trait. The association between atopy-related traits and OA was examined using Mendelian randomization (MR) on summary statistics; we reported inverse-variance weighted (IVW), MR-Egger, maximum likelihood estimation (MLE), weighted median, and weighted mode.There was a significant positive correlation between the genome-wide genetic architecture of asthma and all OA traits. Using the IVW (random effects), there was a significant association between asthma and knee OA (OR=1.04, 95% CI 1.01-1.08, p=0.0169). Using IVW (fixed effects), significant associations were identified between knee OA and allergic disease (OR=1.07, 95% CI 1.01-1.14, p=0.0342), allergic rhinitis (OR=1.07, 95% CI 1.00-1.13, p=0.0368), and asthma (OR=1.04, 95% CI 1.01-1.07, p=0.0139), as well as for OA at any site and asthma (OR=1.02, 95% CI 1.00-1.04, p=0.0166).We found a significant correlation between the overall genetic architecture of asthma and OA, as well as an increased risk of developing OA in patients with genetic variants associated with asthma and allergic rhinitis; predominately, this risk was for the development of knee OA. These results support a causal relationship between asthma and/or allergic rhinitis and knee OA.

  View details for DOI 10.1016/j.joca.2023.11.003

  View details for PubMedID 37951457

• Serum reactivity to citrullinated protein/peptide antigens and left ventricular structure and function in the Multi-Ethnic Study of Atherosclerosis (MESA). PloS one Hughes-Austin, J. M., Katz, R., Majka, D. S., Criqui, M. H., Robinson, W. H., Firestein, G. S., Hundley, W. G., Ix, J. H. 2023; 18 (10): e0291967

  Abstract

  Antibodies to citrullinated protein antigens have been linked to altered left ventricular (LV) structure and function in patients with rheumatoid arthritis (RA). Serum reactivity to several citrullinated protein/peptide antigens has been identified in RA, which are detectable years before RA onset and in individuals who may never develop RA. Among community-living individuals without heart failure (HF) at baseline in the Multi-Ethnic Study of Atherosclerosis (MESA), we investigated associations between serum reactivity to citrullinated protein/peptide antigens, LV mass, LV ejection fraction (LVEF), and incident HF.Among 1232 MESA participants, we measured serum reactivity to 28 different citrullinated proteins/peptides using a multiplex bead-based array. Each antibody was defined as having extremely high reactivity (EHR) if >95th percentile cut-off in MESA. Number of EHR antibody responses to citrullinated protein/peptide antigens were summed for each participant (range 0-28). LV mass(g) and LVEF(%) were measured on cardiac MRI. Associations between EHR antibodies and LV mass and LVEF were evaluated using linear regression. Cox proportional hazards models were used to evaluate associations between EHR antibodies and incident HF during 11 years of follow-up, adjusting for age, gender, race/ethnicity, smoking status, systolic blood pressure, use of anti-hypertensive medications, self-reported arthritis, IL-6, body surface area, and estimated glomerular filtration rate.Mean age was 65±10, 50% were female, 40% were White, 21% were Black, 26% were Hispanic/Latino, and 14% were Chinese. Twenty-seven percent of MESA participants had extremely high reactivity to ≥ 1 citrullinated protein/peptide antigen. In fully adjusted analysis, every additional EHR antibody was significantly associated with 0.1% lower LVEF (95% CI: -0.17%, -0.02%). No association was observed with LV mass (β per additional EHR antibody) = 0.13±0.15 (p = 0.37)). Neither the presence nor number of EHR antibodies was associated with incident HF during follow-up (HR per additional EHR antibody = 1.008 (95% CI: 0.97, 1.05)).Greater number of extremely highly reactive antibodies was associated with lower LVEF, but not with LV mass or incident HF. Thus, serum reactivity to citrullinated protein/peptide antigens was associated with subtle subclinical changes in myocardial contractility, but the significance in relation to clinically apparent HF is uncertain.

  View details for DOI 10.1371/journal.pone.0291967

  View details for PubMedID 37874814

• Does EBV infection matter for MS: EBV mediated molecular mimicry Robinson, W. H. SAGE PUBLICATIONS LTD. 2023: 7

  View details for Web of Science ID 001091311300010

• Anti-citrullinated protein antibodies with multiple specificities ameliorate collagen antibody-induced arthritis in a time-dependent manner. Arthritis & rheumatology (Hoboken, N.J.) Gomez, A. M., Brewer, R. C., Moon, J. S., Acharya, S., Kongpachith, S., Wang, Q., Jahanbani, S., Wong, H. H., Lanz, T. V., Love, Z. Z., Min-Oo, G., Niedziela-Majka, A., Robinson, W. H. 2023

  Abstract

  Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA) and have long been regarded as pathogenic. Despite substantial in vitro evidence supporting this claim, reports investigating the pro-inflammatory effects of ACPAs in animal models of arthritis are rare and include mixed results. Here, we sequenced the plasmablast antibody repertoire of a RA patient and functionally characterized the encoded ACPAs.We expressed ACPAs from the antibody repertoire of a RA patient and characterized their autoantigen specificities on antigen arrays and ELISAs. Binding affinities were estimated by bio-layer interferometry. Select ACPAs (n=9) were tested in the collagen-antibody induced arthritis (CAIA) mouse model, to evaluate their effects on joint inflammation.Recombinant ACPAs bound preferentially, and with high affinity (nM range), to citrullinated (cit) autoantigens (primarily histones and fibrinogen), and to auto-citrullinated peptidylarginine deiminase 4 (PAD4). ACPAs were grouped for in vivo testing based on their predominant cit-antigen specificities. Unexpectedly, injections of recombinant ACPAs significantly reduced paw thickness and arthritis severity in CAIA mice, as compared to isotype-matched control antibodies (p≤0.001). Bone erosion, synovitis, and cartilage damage were also significantly reduced (p≤0.01). This amelioration of CAIA was observed for all the ACPAs tested and was independent of cit-PAD4 and cit-fibrinogen specificities. Further, disease amelioration was more prominent when ACPAs were injected at earlier stages of CAIA than at later phases of the model.Recombinant, patient-derived ACPAs ameliorated CAIA. Their anti-inflammatory effects were more preventative than therapeutic. This study highlights a potential protective role for ACPAs in arthritis.

  View details for DOI 10.1002/art.42679

  View details for PubMedID 37610274

• Anticitrullinated peptide antibody epitope expansion and the HLA DRB1 'shared epitope' are less common in seropositive checkpoint inhibitor-induced inflammatory arthritis than in longstanding rheumatoid arthritis. RMD open Ghosh, N., Reid, P., Aude, C. A., Kirschman, J., Goodman, S., Bykerk, V. P., Lakhanpal, A., Rajesh, D., Chan, K. K., Robinson, W. H., Bass, A. R. 2023; 9 (2)

  Abstract

  Immune checkpoint inhibitors (ICI) can potentially cause ICI-inflammatory arthritis (ICI-IA), which often resembles rheumatoid arthritis (RA). In this study, we examined the degree of anticitrullinated peptide antibodies (ACPA) epitope expansion in CCP+ICI-IA and patients with RA.We used clinical data and serum from ICI-IA and patients with RA with early disease as well as longstanding disease. A custom, bead-based antigen array was used to identify IgG ACPA reactivities to 18 putative RA-associated citrullinated proteins. Hierarchical clustering software was used to create a heatmap to identify ACPA levels. Additionally, HLA DRB1 typing was performed on ICI-IA patients as well as controls of patients treated with ICI that did not develop ICI-IA (ICI controls).Compared to patients with CCP+RA, patients with CCP+ICI-IA were older (p<0.001), less likely to have positive rheumatoid factor (p<0.001) and had a shorter duration of symptoms (p<0.001). There were less ACPA levels and a lower number of distinct ACPA epitopes in the serum of patients with ICI-IA compared with longstanding patients with RA (p<0.001). Among those tested for HLA DRB1, there were no differences in the frequency of the shared epitope between those with ICI-IA and ICI controls.Patients with ICI-IA had lower ACPA titres and targeted fewer ACPA epitopes than longstanding patients with RA, and there were no significant differences in the presence of the shared epitope between those that developed ICI-IA and ICI controls. It remains to be determined if ICI-IA represents an accelerated model of RA pathogenesis with ICI triggering a transition from preclinical to clinical disease.

  View details for DOI 10.1136/rmdopen-2023-003012

  View details for PubMedID 37355249

• Anti-citrullinated protein antibody profiles predict changes in disease activity in patients with rheumatoid arthritis initiating biologics RHEUMATOLOGY Aripova, N., Kremer, J. M., Pappas, D. A., Reed, G., England, B. R., Robinson, B. H., Curtis, J. R., Thiele, G. M., Mikuls, T. R. 2023

  Abstract

  To determine whether an expanded antigen-specific anti-citrullinated protein antibody (ACPA) profile predicts changes in disease activity in patients with rheumatoid arthritis (RA) initiating biologics.The study included participants from a prospective, non-randomized, observational RA cohort. For this sub-study, treatment groups of interest included biologic-naïve initiating anti-TNF, biologic-exposed initiating non-TNF, and biologic-naïve initiating abatacept. ACPAs to 25 citrullinated peptides were measured using banked enrolment serum. Principal component analysis (PCA) was performed and associations of resulting principal component (PC) scores (in quartiles) and anti-CCP3 antibody (≤15, 16-250 or > 250 U/ml) with EULAR (good/moderate/none) treatment response at 6-months were examined using adjusted ordinal regression models.Participants (n = 1092) had a mean age of 57 (±13) years and 79% were women. At 6-months, 68.5% achieved a moderate/good EULAR response. There were 3 PCs that cumulatively explained 70% of variation in ACPA values. In models including the 3 components and anti-CCP3 antibody category, only PC1 and PC2 were associated with treatment response. The highest quartile for PC1 (OR 1.76; 95% CI 1.22-2.53) and for PC2 (OR 1.74; 95% CI 1.23-2.46) were associated with treatment response after multivariable adjustment. There was no evidence of interaction between PCs and treatment group in EULAR responses (p-for-interaction >0.1).An expanded ACPA profile appears to be more strongly associated with biologic treatment response in RA than commercially available anti-CCP3 antibody levels. However, further enhancements to PCA will be needed to effectively prioritize between different biologics available for the treatment of RA.

  View details for DOI 10.1093/rheumatology/kead260

  View details for Web of Science ID 001011173700001

  View details for PubMedID 37252826

• Single-cell sequencing of PBMC characterizes a distinct inflammatory monocyte subset in Lyme disease patients with Post-Treatment Lyme Disease Syndrome (PTLD) Chen, X., Zhou, X., Ji, X., Miranda, M., Rebman, A., Kathuria, K., Robinson, W. H., Aucott, J., Davis, M. M. AMER ASSOC IMMUNOLOGISTS. 2023

  View details for DOI 10.4049/jimmunol.210.Supp.155.18

  View details for Web of Science ID 001106506501370

• Rheumatoid arthritis associated with a cytotoxic cell leukemia is characterized by enhanced reactivity to citrullinated antigens Ananth, K., Gomez-Banuelos, E., Trejo-Zambrano, I., Kirschmann, J., Moosic, K. B., Wang, H., Feith, D. J., Robinson, W. H., Andrade, F., Loughran, T. P., Darrah, E. AMER ASSOC IMMUNOLOGISTS. 2023

  View details for Web of Science ID 001106506503254

• Clonally Expanded Cytotoxic CD8<SUP>+</SUP> T cells Target Citrullinated Antigens In ACPA<SUP>+</SUP> Rheumatoid Arthritis Moon, J., Younis, S., Sharpe, O., Rao, N. L., Carman, J. A., James, E. A., Buckner, J. H., Deane, K. D., Holers, V., Donlin, L. T., Davis, M. M., Robinson, W. H. AMER ASSOC IMMUNOLOGISTS. 2023

  View details for Web of Science ID 001106506500443

• Increased risk of osteoarthritis in patients with atopic disease. Annals of the rheumatic diseases Baker, M. C., Sheth, K., Lu, R., Lu, D., von Kaeppler, E. P., Bhat, A., Felson, D. T., Robinson, W. H. 2023

  Abstract

  To determine the incidence of osteoarthrits (OA) in patients with atopic disease compared with matched non-exposed patients.We conducted a retrospective cohort study with propensity score matching using claims data from Optum's de-identified Clinformatics Data Mart (CDM) (January 2003 to June 2019) and electronic health record data from the Stanford Research Repository (STARR) (January 2010 to December 2020). We included adult patients without pre-existing OA or inflammatory arthritis who were exposed to atopic disease or who were non-exposed. The primary outcome was the development of incident OA.In Optum CDM, we identified 117 346 exposed patients with asthma or atopic dermatitis (mean age 52 years; 60% female) and 1 247 196 non-exposed patients (mean age 50 years; 48% female). After propensity score matching (n=1 09 899 per group), OA incidence was higher in patients with asthma or atopic dermatitis (26.9 per 1000 person-years) compared with non-exposed patients (19.1 per 1000 person-years), with an adjusted odds ratio (aOR) of 1.58 (95% CI 1.55 to 1.62) for developing OA. This effect was even more pronounced in patients with both asthma and atopic dermatitis compared with non-exposed patients (aOR=2.15; 95% CI 1.93 to 2.39) and in patients with asthma compared with patients with chronic obstructive pulmonary disease (aOR=1.83; 95% CI 1.73 to 1.95). We replicated our results in an independent dataset (STARR), which provided the added richness of body mass index data. The aOR of developing OA in patients with asthma or atopic dermatitis versus non-exposed patients in STARR was 1.42 (95% CI 1.36 to 1.48).This study demonstrates an increased incidence of OA in patients with atopic disease. Future interventional studies may consider targeting allergic pathways for the prevention or treatment of OA.

  View details for DOI 10.1136/ard-2022-223640

  View details for PubMedID 36987654

• FAT10 Induces cancer cell migration by stabilizing phosphorylated ABI3/NESH. Animal cells and systems Um, H., Jeong, H., Lee, B., Kim, Y., Lee, J., Roh, J. S., Lee, S. G., Park, H. R., Robinson, W. H., Sohn, D. H. 2023; 27 (1): 53-60

  Abstract

  The WAVE regulatory complex (WRC) is involved in various cellular processes by regulating actin polymerization. The dysregulation of WRC components is associated with cancer development. ABI family member 3 (ABI3)/new molecule including SH3 (NESH) is one of the WRC components and it has been reported that ABI3 phosphorylation can affect WRC function. Although several residues of ABI3 have been reported to be possible phosphorylation sites, it is still unclear which residues are important for the function of ABI3. Furthermore, it is unclear how the phosphorylated form of ABI3 is regulated. Here, we demonstrate that ABI3 is stabilized by its interaction with human leukocyte antigen-F adjacent transcript 10 (FAT10). Using phospho-dead or phospho-mimetic mutants of ABI3, we showed that serine 213 and 216 are important phosphorylation sites of ABI3. In particular, FAT10 has a higher affinity for the phosphorylated form of ABI3 than the non-phosphorylated form, and it stabilizes the phosphorylated form more than the non-phosphorylated form through this differential affinity. The interaction between FAT10 and the phosphorylated form of ABI3 promoted cancer cell migration. Therefore, our results suggest that FAT10 stabilizes the phosphorylated form of ABI3, which may lead to WRC activation, thereby promoting cancer cell migration.

  View details for DOI 10.1080/19768354.2023.2186486

  View details for PubMedID 36926204

  View details for PubMedCentralID PMC10013321

• Dysfunction in parkin aggravates inflammatory bone erosion by reinforcing osteoclast activity. Cell & bioscience Kim, E., Kim, J., Kim, Y., Choi, B., Sohn, D. H., Park, S., Chung, Y., Kim, Y., Robinson, W. H., Kim, Y., Chang, E. 2023; 13 (1): 48

  Abstract

  BACKGROUND: Parkin dysfunction associated with the progression of parkinsonism contributes to a progressive systemic skeletal disease characterized by low bone mineral density. However, the role of parkin in bone remodeling has not yet been elucidated in detail.RESULT: We observed that decreased parkin in monocytes is linked to osteoclastic bone-resorbing activity. siRNA-mediated knockdown of parkin significantly enhanced the bone-resorbing activity of osteoclasts (OCs) on dentin without any changes in osteoblast differentiation. Moreover, Parkin-deficient mice exhibited an osteoporotic phenotype with a lower bone volume accompanied by increased OC-mediated bone-resorbing capacity displaying increased acetylation of alpha-tubulin compared to wild-type (WT) mice. Notably, compared to WT mice, the Parkin-deficient mice displayed increased susceptibility to inflammatory arthritis, reflected by a higher arthritis score and a marked bone loss after arthritis induction using K/BxN serum transfer, but not ovariectomy-induced bone loss. Intriguingly, parkin colocalized with microtubules and parkin-depleted-osteoclast precursor cells (Parkin-/- OCPs) displayed augmented ERK-dependent acetylation of alpha-tubulin due to failure of interaction with histone deacetylase 6 (HDAC6), which was promoted by IL-1beta signaling. The ectopic expression of parkin in Parkin-/- OCPs limited the increase in dentin resorption induced by IL-1beta, accompanied by the reduced acetylation of alpha-tubulin and diminished cathepsin K activity.CONCLUSION: These results indicate that a deficiency in the function of parkin caused by a decrease in parkin expression in OCPs under the inflammatory condition may enhance inflammatory bone erosion by altering microtubule dynamics to maintain OC activity.

  View details for DOI 10.1186/s13578-023-00973-0

  View details for PubMedID 36882866

• Machine learning identification of thresholds to discriminate osteoarthritis and rheumatoid arthritis synovial inflammation. Arthritis research & therapy Mehta, B., Goodman, S., DiCarlo, E., Jannat-Khah, D., Gibbons, J. A., Otero, M., Donlin, L., Pannellini, T., Robinson, W. H., Sculco, P., Figgie, M., Rodriguez, J., Kirschmann, J. M., Thompson, J., Slater, D., Frezza, D., Xu, Z., Wang, F., Orange, D. E. 2023; 25 (1): 31

  Abstract

  We sought to identify features that distinguish osteoarthritis (OA) and rheumatoid arthritis (RA) hematoxylin and eosin (H&E)-stained synovial tissue samples.We compared fourteen pathologist-scored histology features and computer vision-quantified cell density (147 OA and 60 RA patients) in H&E-stained synovial tissue samples from total knee replacement (TKR) explants. A random forest model was trained using disease state (OA vs RA) as a classifier and histology features and/or computer vision-quantified cell density as inputs.Synovium from OA patients had increased mast cells and fibrosis (p < 0.001), while synovium from RA patients exhibited increased lymphocytic inflammation, lining hyperplasia, neutrophils, detritus, plasma cells, binucleate plasma cells, sub-lining giant cells, fibrin (all p < 0.001), Russell bodies (p = 0.019), and synovial lining giant cells (p = 0.003). Fourteen pathologist-scored features allowed for discrimination between OA and RA, producing a micro-averaged area under the receiver operating curve (micro-AUC) of 0.85±0.06. This discriminatory ability was comparable to that of computer vision cell density alone (micro-AUC = 0.87±0.04). Combining the pathologist scores with the cell density metric improved the discriminatory power of the model (micro-AUC = 0.92±0.06). The optimal cell density threshold to distinguish OA from RA synovium was 3400 cells/mm2, which yielded a sensitivity of 0.82 and specificity of 0.82.H&E-stained images of TKR explant synovium can be correctly classified as OA or RA in 82% of samples. Cell density greater than 3400 cells/mm2 and the presence of mast cells and fibrosis are the most important features for making this distinction.

  View details for DOI 10.1186/s13075-023-03008-8

  View details for PubMedID 36864474

  View details for PubMedCentralID PMC9979511

• Development of Osteoarthritis in Adults With Type 2 Diabetes Treated With Metformin vs a Sulfonylurea. JAMA network open Baker, M. C., Sheth, K., Liu, Y., Lu, D., Lu, R., Robinson, W. H. 2023; 6 (3): e233646

  Abstract

  Importance: Metformin may have a protective association against developing osteoarthritis (OA), but robust epidemiological data are lacking.Objective: To determine the risk of OA and joint replacement in individuals with type 2 diabetes treated with metformin compared with a sulfonylurea.Design, Setting, and Participants: This retrospective cohort study used claims data from the Optum deidentified Clinformatics Data Mart Database between December 2003 and December 2019. Participants included individuals aged 40 years or older with at least 1 year of continuous enrollment and type 2 diabetes. Individuals with type 1 diabetes or a prior diagnosis of OA, inflammatory arthritis, or joint replacement were excluded. Time-conditional propensity score matching was conducted using age, sex, race, Charlson comorbidity score, and treatment duration to create a prevalent new-user cohort. Data were analyzed from April to December 2021.Exposures: Treatment with metformin or a sulfonylurea.Main Outcomes and Measures: The outcomes of interest were incident OA and joint replacement. Cox proportional hazard models were used to calculate adjusted hazard ratios (aHRs) of incident OA and joint replacement. In a sensitivity analysis, individuals only ever treated with metformin were compared with individuals only ever treated with a sulfonylurea, allowing for longer-term follow up of the outcome (even after stopping the medication of interest).Results: After time-conditional propensity score matching, the metformin and control groups each included 20 937 individuals (mean [SD] age 62.0 [11.5] years; 24 379 [58.2%] males). In the adjusted analysis, the risk of developing OA was reduced by 24% for individuals treated with metformin compared with a sulfonylurea (aHR, 0.76; 95% CI, 0.68-0.85; P<.001), but there was no significant difference for risk of joint replacement (aHR, 0.80; 95% CI, 0.50-1.27; P=.34). In the sensitivity analysis, the risk of developing OA remained lower in individuals treated with metformin compared with a sulfonylurea (aHR, 0.77; 95% CI, 0.65-0.90; P<.001) and the risk of joint replacement remained not statistically significant (aHR, 1.04; 95% CI, 0.60-1.82; P=.89).Conclusions and Relevance: In this cohort study of individuals with diabetes, metformin treatment was associated with a significant reduction in the risk of developing OA compared with sulfonylurea treatment. These results further support preclinical and observational data that suggest metformin may have a protective association against the development of OA; future interventional studies with metformin for the treatment or prevention of OA should be considered.

  View details for DOI 10.1001/jamanetworkopen.2023.3646

  View details for PubMedID 36939700

• Incidence of Interstitial Lung Disease in Patients With Rheumatoid Arthritis Treated With Biologic and Targeted Synthetic Disease-Modifying Antirheumatic Drugs. JAMA network open Baker, M. C., Liu, Y., Lu, R., Lin, J., Melehani, J., Robinson, W. H. 2023; 6 (3): e233640

  Abstract

  Importance: Current data are lacking regarding the risk of biologic and targeted synthetic disease-modifying antirheumatic drug (b/tsDMARD) use on the development of interstitial lung disease (ILD) in patients with rheumatoid arthritis (RA).Objective: To determine the risk of developing ILD in patients with RA undergoing treatment with different b/tsDMARDs.Design, Setting, and Participants: Retrospective cohort study using claims data from the Optum Clinformatics Data Mart between December 2003 and December 2019. Adult patients with RA, 1 year or more of continuous enrollment, treatment with a b/tsDMARD of interest, and without preexisting ILD were included. Data were analyzed from October 2021 to April 2022.Exposures: New administration of adalimumab, abatacept, rituximab, tocilizumab, or tofacitinib.Main Outcomes and Measures: Crude incidence rates (IRs) for the development of ILD were calculated. The risk of ILD across different b/tsDMARDs was compared using Cox-regression models. A sensitivity analysis using a prevalent new-user cohort design compared patients treated with tofacitinib and adalimumab.Results: A total of 28 559 patients with RA (mean [SD] age 55.6 [13.7] years; 22 158 female [78%]) were treated with adalimumab (13 326 patients), abatacept (5676 patients), rituximab (5444 patients), tocilizumab (2548 patients), or tofacitinib (1565 patients). Crude IRs per 1000 person-years for ILD were 3.43 (95% CI 2.85-4.09) for adalimumab, 4.46 (95% CI 3.44-5.70) for abatacept, 6.15 (95% CI 4.76-7.84) for rituximab, 5.05 (95% CI 3.47-7.12) for tocilizumab, and 1.47 (95% CI 0.54-3.27) for tofacitinib. After multiple adjustments, compared with patients treated with adalimumab, patients treated with tofacitinib had a lower risk of ILD (adjusted hazard ratio [aHR] 0.31; 95% CI, 0.12-0.78; P=.009). In a prevalent new-user cohort analysis, patients treated with tofacitinib had 68% reduced risk of ILD compared with adalimumab (aHR 0.32; 95% CI 0.13-0.82; P<.001). In an adjusted model, there was a 69% reduced risk of ILD in patients treated with tofacitinib compared with patients treated with adalimumab.Conclusions and Relevance: In this retrospective cohort of patients with RA, patients treated with tofacitinib had the lowest incidence of ILD compared with patients treated with all bDMARDs evaluated, and patients treated with tofacitinib had a reduced risk of ILD compared with patients treated with adalimumab after adjusting for important covariates. Additional prospective studies are needed to better understand the role tofacitinib may play in preventing ILD in patients with RA. These results, while significant, should be interpreted with caution given the fairly small sample size of the tofacitinib group.

  View details for DOI 10.1001/jamanetworkopen.2023.3640

  View details for PubMedID 36939701

• Oral mucosal breaks trigger anti-citrullinated bacterial and human protein antibody responses in rheumatoid arthritis. Science translational medicine Brewer, R. C., Lanz, T. V., Hale, C. R., Sepich-Poore, G. D., Martino, C., Swafford, A. D., Carroll, T. S., Kongpachith, S., Blum, L. K., Elliott, S. E., Blachere, N. E., Parveen, S., Fak, J., Yao, V., Troyanskaya, O., Frank, M. O., Bloom, M. S., Jahanbani, S., Gomez, A. M., Iyer, R., Ramadoss, N. S., Sharpe, O., Chandrasekaran, S., Kelmenson, L. B., Wang, Q., Wong, H., Torres, H. L., Wiesen, M., Graves, D. T., Deane, K. D., Holers, V. M., Knight, R., Darnell, R. B., Robinson, W. H., Orange, D. E. 2023; 15 (684): eabq8476

  Abstract

  Periodontal disease is more common in individuals with rheumatoid arthritis (RA) who have detectable anti-citrullinated protein antibodies (ACPAs), implicating oral mucosal inflammation in RA pathogenesis. Here, we performed paired analysis of human and bacterial transcriptomics in longitudinal blood samples from RA patients. We found that patients with RA and periodontal disease experienced repeated oral bacteremias associated with transcriptional signatures of ISG15+HLADRhi and CD48highS100A2pos monocytes, recently identified in inflamed RA synovia and blood of those with RA flares. The oral bacteria observed transiently in blood were broadly citrullinated in the mouth, and their in situ citrullinated epitopes were targeted by extensively somatically hypermutated ACPAs encoded by RA blood plasmablasts. Together, these results suggest that (i) periodontal disease results in repeated breaches of the oral mucosa that release citrullinated oral bacteria into circulation, which (ii) activate inflammatory monocyte subsets that are observed in inflamed RA synovia and blood of RA patients with flares and (iii) activate ACPA B cells, thereby promoting affinity maturation and epitope spreading to citrullinated human antigens.

  View details for DOI 10.1126/scitranslmed.abq8476

  View details for PubMedID 36812347

• Serum antibodies to periodontal pathogens prior to rheumatoid arthritis diagnosis: A case-control study. Seminars in arthritis and rheumatism Lee, J. A., Mikuls, T. R., Deane, K. D., Sayles, H. R., Thiele, G. M., Edison, J. D., Wagner, B. D., Feser, M. L., Moss, L. K., Kelmenson, L. B., Robinson, W. H., Payne, J. B. 2023; 59: 152176

  Abstract

  OBJECTIVES: 1) To quantify the association between anti-Porphyromonas gingivalis serum antibody concentrations and the risk of developing rheumatoid arthritis (RA), and 2) to quantify the associations among RA cases between anti-P. gingivalis serum antibody concentrations and RA-specific autoantibodies. Additional anti-bacterial antibodies evaluated included anti-Fusobacterium nucleatum and anti-Prevotella intermedia.METHODS: Serum samples were acquired pre- and post- RA diagnosis from the U.S. Department of Defense Serum Repository (n=214 cases, 210 matched controls). Using separate mixed-models, the timing of elevations of anti-P. gingivalis, anti-P. intermedia, and anti-F. nucleatum antibody concentrations relative to RA diagnosis were compared in RA cases versus controls. Associations were determined between serum anti-CCP2, ACPA fine specificities (vimentin, histone, and alpha-enolase), and IgA, IgG, and IgM RF in pre-RA diagnosis samples and anti-bacterial antibodies using mixed-effects linear regression models.RESULTS: No compelling evidence of case-control divergence in serum anti-P. gingivalis, anti-F. nucleatum, and anti-P. intermedia was observed. Among RA cases, including all pre-diagnosis serum samples, anti-P. intermedia was significantly positively associated with anti-CCP2, ACPA fine specificities targeting vimentin, histone, alpha-enolase, and IgA RF (p<0.001), IgG RF (p=0.049), and IgM RF (p=0.004), while anti-P. gingivalis and anti-F. nucleatum were not.CONCLUSIONS: No longitudinal elevations of anti-bacterial serum antibody concentrations were observed in RA patients prior to RA diagnosis compared to controls. However, anti-P. intermedia displayed significant associations with RA autoantibody concentrations prior to RA diagnosis, suggesting a potential role of this organism in progression towards clinically-detectable RA.

  View details for DOI 10.1016/j.semarthrit.2023.152176

  View details for PubMedID 36812865

• Autoantibodies against citrullinated and native proteins and prediction of rheumatoid arthritis-associated interstitial lung disease: A nested case-control study. The Lancet. Rheumatology Kronzer, V. L., Hayashi, K., Yoshida, K., Davis, J. M., McDermott, G. C., Huang, W., Dellaripa, P. F., Cui, J., Feathers, V., Gill, R. R., Hatabu, H., Nishino, M., Blaustein, R., Crowson, C. S., Robinson, W. H., Sokolove, J., Liao, K. P., Weinblatt, M. E., Shadick, N. A., Doyle, T. J., Sparks, J. A. 2023; 5 (2): e77-e87

  Abstract

  Background: To identify fine specificity anti-citrullinated protein antibodies (ACPA) associated with incident rheumatoid arthritis-associated interstitial lung disease (RA-ILD).Methods: This nested case-control study within the Brigham RA Sequential Study matched incident RA-ILD cases to RA-noILD controls on time of blood collection, age, sex, RA duration, and rheumatoid factor status. A multiplex assay measured ACPA and anti-native protein antibodies from stored serum prior to RA-ILD onset. Logistic regression models calculated odds ratios (OR) with 95% confidence intervals (CI) for RA-ILD, adjusting for prospectively-collected covariates. We estimated optimism-corrected area under the curves (AUC) using internal validation. Model coefficients generated a risk score for RA-ILD.Findings: We analyzed 84 incident RA-ILD cases (mean age 67 years, 77% female, 90% White) and 233 RA-noILD controls (mean age 66 years, 80% female, 94% White). We identified six fine specificity antibodies that were associated with RA-ILD. The antibody isotypes and targeted proteins were: IgA2 to citrullinated histone 4 (OR 0.08 per log-transformed unit, 95% CI 0.03-0.22), IgA2 to citrullinated histone 2A (OR 4.03, 95% CI 2.03-8.00), IgG to cyclic citrullinated filaggrin (OR 3.47, 95% CI 1.71-7.01), IgA2 to native cyclic histone 2A (OR 5.52, 95% CI 2.38-12.78), IgA2 to native histone 2A (OR 4.60, 95% CI 2.18-9.74), and IgG to native cyclic filaggrin (OR 2.53, 95% CI 1.47-4.34). These six antibodies predicted RA-ILD risk better than all clinical factors combined (optimism-corrected AUC=0·84 versus 0·73). We developed a risk score for RA-ILD combining these antibodies with the clinical factors (smoking, disease activity, glucocorticoid use, obesity). At 50% predicted RA-ILD probability, the risk scores both without (score=2·6) and with (score=5·9) biomarkers achieved specificity ≥93% for RA-ILD.Interpretation: Specific ACPA and anti-native protein antibodies improve RA-ILD prediction. These findings implicate synovial protein antibodies in the pathogenesis of RA-ILD and suggest clinical utility in predicting RA-ILD once validated in external studies.Funding: National Institutes of Health.

  View details for DOI 10.1016/s2665-9913(22)00380-0

  View details for PubMedID 36874209

• Autoantibodies against citrullinated and native proteins and prediction of rheumatoid arthritis-associated interstitial lung disease: a nested case-control study LANCET RHEUMATOLOGY Kronzer, V. L., Hayashi, K., Yoshida, K., Davis, J. M., McDermott, G. C., Huang, W., Dellaripa, P. F., Cui, J., Feathers, V., Gill, R. R., Hatabu, H., Nishino, M., Blaustein, R., Crowson, C. S., Robinson, W. H., Sokolove, J., Liao, K. P., Weinblatt, M. E., Shadick, N. A., Doyle, T. J., Sparks, J. A. 2023; 5 (2): E77-E87

  View details for Web of Science ID 001031413300001

• Cytotoxic CD8+ T cells target citrullinated antigens in rheumatoid arthritis. Nature communications Moon, J. S., Younis, S., Ramadoss, N. S., Iyer, R., Sheth, K., Sharpe, O., Rao, N. L., Becart, S., Carman, J. A., James, E. A., Buckner, J. H., Deane, K. D., Holers, V. M., Goodman, S. M., Donlin, L. T., Davis, M. M., Robinson, W. H. 2023; 14 (1): 319

  Abstract

  The immune mechanisms that mediate synovitis and joint destruction in rheumatoid arthritis (RA) remain poorly defined. Although increased levels of CD8+ T cells have been described in RA, their function in pathogenesis remains unclear. Here we perform single cell transcriptome and T cell receptor (TCR) sequencing of CD8+ T cells derived from anti-citrullinated protein antibodies (ACPA)+ RA blood. We identify GZMB+CD8+ subpopulations containing large clonal lineage expansions that express cytotoxic and tissue homing transcriptional programs, while a GZMK+CD8+ memory subpopulation comprises smaller clonal expansions that express effector T cell transcriptional programs. We demonstrate RA citrullinated autoantigens presented by MHC class I activate RA blood-derived GZMB+CD8+ T cells to expand, express cytotoxic mediators, and mediate killing of target cells. We also demonstrate that these clonally expanded GZMB+CD8+ cells are present in RA synovium. These findings suggest that cytotoxic CD8+ T cells targeting citrullinated antigens contribute to synovitis and joint tissue destruction in ACPA+ RA.

  View details for DOI 10.1038/s41467-022-35264-8

  View details for PubMedID 36658110

  View details for PubMedCentralID PMC9852471

• Roadmap for understanding mechanisms on how Epstein-Barr virus triggers multiple sclerosis and for translating these discoveries in clinical trials. Clinical & translational immunology Lanz, T. V., Robinson, W. H., Ho, P. P., Steinman, L. 2023; 12 (2): e1438

  Abstract

  Here, we offer a roadmap for what might be studied next in understanding how EBV triggers MS. We focus on two areas: The first area concerns the molecular mechanisms underlying how clonal antibody in the CSF emanates in widespread molecular mimicry to key antigens in the nervous system including GlialCAM, a protein associated with chloride channels. A second and equally high priority in the roadmap concerns various therapeutic approaches that are related to blocking the mechanisms whereby EBV triggers MS. Therapies deserving of attention include clinical trials with antivirals and the development of 'inverse' vaccines based on nucleic acid technologies to control or to eradicate the consequences of EBV infection. High enthusiasm is given to continuation of ongoing clinical trials of cellular adoptive therapy to attack EBV-infected cells. Clinical trials of vaccines to EBV are another area deserving attention. These suggested topics involving research on mechanism, and the design, implementation and performance of well-designed trials are not intended to be an exhaustive list. We have splendid tools available to our community of medical scientists to tackle how EBV triggers MS and then to perhaps change the world with new therapies to potentially eradicate MS, as we have done with nearly complete success for poliomyelitis.

  View details for DOI 10.1002/cti2.1438

  View details for PubMedID 36815946

• Activation of a Latent Epitope Causing Differential Binding of Anti-Neutrophil Cytoplasmic Antibodies to Proteinase 3. Arthritis & rheumatology (Hoboken, N.J.) Moura, M. C., Thompson, G. E., Nelson, D. R., Fussner, L. A., Hummel, A. M., Jenne, D. E., Emerling, D., Fervenza, F. C., Kallenberg, C. G., Langford, C. A., McCune, W. J., Merkel, P. A., Monach, P. A., Seo, P., Spiera, R. F., St Clair, E. W., Ytterberg, S. R., Stone, J. H., Robinson, W. H., Specks, U., WGET and RAVE-ITN Research Groups 2022

  Abstract

  OBJECTIVE: Proteinase 3 (PR3) is the major antigen for anti-neutrophil cytoplasmic antibodies (ANCAs) in the systemic autoimmune vasculitis, granulomatosis with polyangiitis (GPA). PR3 anti-neutrophil cytoplasmic antibodies (PR3-ANCAs) recognize different epitopes on PR3. We aimed to study the effect of mutations on PR3 antigenicity.METHODS: The recombinant PR3 variants, iPR3 (clinically used to detect PR3-ANCAs) and iHm5 (containing three point mutations in Epitope 1&5 generated for epitope mapping studies, immunoassays and serum samples from patients enrolled in ANCA-associated vasculitis (AAV) trials were used to screen the differential PR3-ANCA binding. A patient-derived monoclonal ANCA (moANCA518) that selectively binds to iHm5 within the mutation-free Epitope 3 and distant from the point mutations of iHm5 was used as a gauge of remote epitope activation. Selective binding was determined by inhibition experiments.RESULTS: Rather than reduced binding of PR3-ANCAs to iHm5, we found substantially increased binding of the majority of PR3-ANCAs to iHm5 compared with iPR3. This differential binding of PR3-ANCA to iHm5 is similar to the selective moANCA518 binding to iHm5. Binding of iPR3 to monoclonal antibody MCPR3-2 also induced recognition by moANCA518.CONCLUSION: The preferential binding of PR3-ANCAs from patients like the selective binding of moANCA518 to iHm5 is conferred by increased antigenicity of Epitope 3 on iHm5. This can also be induced on iPR3 when captured by monoclonal antibody MCPR-2. This previously unrecognized characteristic of PR3-ANCA interactions with its target antigen has implications for studying antibody-mediated autoimmune diseases, understanding of variable performance characteristics of immunoassays and design of potential novel treatment approaches. This article is protected by copyright. All rights reserved.

  View details for DOI 10.1002/art.42418

  View details for PubMedID 36515151

• Increased macrophage phagocytic activity with TLR9 agonist conjugation of an anti- Borrelia burgdorferi monoclonal antibody. Clinical immunology (Orlando, Fla.) Jahanbani, S., Hansen, P. S., Blum, L. K., Bastounis, E. E., Ramadoss, N. S., Pandrala, M., Kirschmann, J. M., Blacker, G. S., Love, Z. Z., Weissman, I. L., Nemati, F., Tal, M. C., Robinson, W. H. 2022: 109180

  Abstract

  Borrelia burgdorferi (Bb) infection causes Lyme disease, for which there is need for more effective therapies. Here, we sequenced the antibody repertoire of plasmablasts in Bb-infected humans. We expressed recombinant monoclonal antibodies (mAbs) representing the identified plasmablast clonal families, and identified their binding specificities. Our recombinant anti-Bb mAbs exhibit a range of activity in mediating macrophage phagocytosis of Bb. To determine if we could increase the macrophage phagocytosis-promoting activity of our anti-Bb mAbs, we generated a TLR9-agonist CpG-oligo-conjugated anti-BmpA mAb. We demonstrated that our CpG-conjugated anti-BmpA mAb exhibited increased peak Bb phagocytosis at 12-24 h, and sustained macrophage phagocytosis over 60+ hrs. Further, our CpG-conjugated anti-BmpA mAb induced macrophages to exhibit a sustained activation morphology. Our findings demonstrate the potential for TLR9-agonist CpG-oligo conjugates to enhance mAb-mediated clearance of Bb, and this approach might also enhance the activity of other anti-microbial mAbs.

  View details for DOI 10.1016/j.clim.2022.109180

  View details for PubMedID 36396013

• Clonal IgA and IgG autoantibodies from individuals at risk for rheumatoid arthritis identify an arthritogenic strain of Subdoligranulum SCIENCE TRANSLATIONAL MEDICINE Chriswell, M. E., Lefferts, A. R., Clay, M. R., Hsu, A., Seifert, J., Feser, M. L., Rims, C., Bloom, M. S., Bemis, E. A., Liu, S., Maerz, M. D., Frank, D. N., Demoruelle, M., Deane, K. D., James, E. A., Buckner, J. H., Robinson, W. H., Holers, V., Kuhn, K. A. 2022; 14 (668)

  View details for Web of Science ID 000909210700001

• Clonal IgA and IgG autoantibodies from individuals at risk for rheumatoid arthritis identify an arthritogenic strain of Subdoligranulum. Science translational medicine Chriswell, M. E., Lefferts, A. R., Clay, M. R., Hsu, A. R., Seifert, J., Feser, M. L., Rims, C., Bloom, M. S., Bemis, E. A., Liu, S., Maerz, M. D., Frank, D. N., Demoruelle, M. K., Deane, K. D., James, E. A., Buckner, J. H., Robinson, W. H., Holers, V. M., Kuhn, K. A. 2022; 14 (668): eabn5166

  Abstract

  The mucosal origins hypothesis of rheumatoid arthritis (RA) proposes a central role for mucosal immune responses in the initiation or perpetuation of the systemic autoimmunity that occurs with disease. However, the connection between the mucosa and systemic autoimmunity in RA remains unclear. Using dual immunoglobulin A (IgA) and IgG family plasmablast-derived monoclonal autoantibodies obtained from peripheral blood of individuals at risk for RA, we identified cross-reactivity between RA-relevant autoantigens and bacterial taxa in the closely related families Lachnospiraceae and Ruminococcaceae. After generating bacterial isolates within the Lachnospiraceae/Ruminococcaceae genus Subdoligranulum from the feces of an individual, we confirmed monoclonal antibody binding and CD4+ T cell activation in individuals with RA compared to control individuals. In addition, when Subdoligranulum isolate 7 but not isolate 1 colonized germ-free mice, it stimulated TH17 cell expansion, serum RA-relevant IgG autoantibodies, and joint swelling reminiscent of early RA, with histopathology characterized by antibody deposition and complement activation. Systemic immune responses were likely due to mucosal invasion along with the generation of colon-isolated lymphoid follicles driving increased fecal and serum IgA by isolate 7, because B and CD4+ T cell depletion not only halted intestinal immune responses but also eliminated detectable clinical disease. In aggregate, these findings demonstrate a mechanism of RA pathogenesis through which a specific intestinal strain of bacteria can drive systemic autoantibody generation and joint-centered antibody deposition and immune activation.

  View details for DOI 10.1126/scitranslmed.abn5166

  View details for PubMedID 36288282

• Myeloablative autologous haematopoietic stem cell transplantation resets the B cell repertoire to a more naïve state in patients with systemic sclerosis. Annals of the rheumatic diseases Adamska, J. Z., Zia, A., Bloom, M. S., Crofford, L. J., Furst, D. E., Goldmuntz, E., Keyes-Elstein, L., Mayes, M. D., McSweeney, P., Nash, R. A., Pinckney, A., Welch, B., Love, Z. Z., Sullivan, K. M., Robinson, W. 2022

  Abstract

  Myeloablative autologous haematopoietic stem cell transplant (HSCT) was recently demonstrated to provide significant benefit over cyclophosphamide (CYC) in the treatment of diffuse cutaneous systemic sclerosis (dcSSc) in the Scleroderma: Cyclophosphamide or Transplantation (SCOT) trial. As dysregulation of the B cell compartment has previously been described in dcSSc, we sought to gain insight into the effects of myeloablative autologous HSCT as compared with CYC.We sequenced the peripheral blood immunoglobulin heavy chain (IGH) repertoires in patients with dcSSc enrolled in the SCOT trial.Myeloablative autologous HSCT was associated with a sustained increase in IgM isotype antibodies bearing a low mutation rate. Clonal expression was reduced in IGH repertoires following myeloablative autologous HSCT. Additionally, we identified a underusage of immunoglobulin heavy chain V gene 5-51 in patients with dcSSc, and usage normalised following myeloablative autologous HSCT but not CYC treatment.Together, these findings suggest that myeloablative autologous HSCT resets the IGH repertoire to a more naïve state characterised by IgM-expressing B cells, providing a possible mechanism for the elimination of pathogenic B cells that may contribute to the benefit of HSCT over CYC in the treatment of dcSSc.

  View details for DOI 10.1136/ard-2021-221925

  View details for PubMedID 36241361

• RNA-seq characterization of histamine-releasing mast cells as potential therapeutic target of osteoarthritis. Clinical immunology (Orlando, Fla.) Zhao, X., Younis, S., Shi, H., Hu, S., Zia, A., Wong, H. H., Elliott, E. E., Chang, T., Bloom, M. S., Zhang, W., Liu, X., Lanz, T. V., Sharpe, O., Love, Z. Z., Wang, Q., Robinson, W. H. 2022: 109117

  Abstract

  OBJECTIVE: Mast cells in the osteoarthritis (OA) synovium correlate with disease severity. This study aimed to further elucidate the role of mast cells in OA by RNA-Seq analysis and pharmacological blockade of the activity of histamine, a key mast cell mediator, in murine OA.METHODS: We examined OA synovial tissues and fluids by flow cytometry, immunostaining, single-cell and bulk RNA-Seq, qPCR, and ELISA. Cetirizine, a histamine H1 receptor (H1R) antagonist, was used to treat the destabilization of the medial meniscus (DMM) mouse model of OA.RESULTS: Flow cytometry and immunohistology analysis of OA synovial cells revealed KIT+ FcepsilonRI+ and TPSAB1+ mast cells. Single-cell RNA-Seq of OA synovial cells identified the expression of prototypical mast cell markers KIT, TPSAB1, CPA3 and HDC, as well as distinctive markers HPGD, CAVIN2, IL1RL1, PRG2, and CKLF, confirmed by bulk RNA-Seq and qPCR. A mast cell prototypical marker expression score classified 40 OA patients into three synovial pathotypes: mast cell-high, -medium, and -low. Additionally, we detected mast cell mediators including histamine, tryptase AB1, CPA3, PRG2, CAVIN2, and CKLF in OA synovial fluids. Elevated H1R expression was detected in human OA synovium, and treatment of mice with the H1 receptor antagonist cetirizine reduced the severity and OA-related mediators in DMM.CONCLUSION: Based on differential expression of prototypical and distinct mast cell markers, human OA joints can be stratified into mast cell-high, -medium, and -low synovial tissue pathotypes. Pharmacologic blockade of histamine activity holds the potential to improve OA disease outcome.

  View details for DOI 10.1016/j.clim.2022.109117

  View details for PubMedID 36109004

• Single-Cell Characterization of the TCR Repertoire Across Tissue and Blood in Rheumatoid Arthritis Dunlap, G., Wagner, A., Meednu, N., Jonsson, A., Zhang, F., Wei, K., Utz, P., Robinson, W., Maecker, H., James, J., Guthridge, J., Bridges, S., Donlin, L., Goodman, S., DiCarlo, E., Bykerk, V., Ritchlin, C., Tabechian, D., Lederer, J., Gravallese, E., McGeachy, M., Firestein, G. S., Gregersen, P., Horowitz, D., Boyle, D., Geraldino-Pardilla, L., Perlman, H., Mandelin, A., Bathon, J., Hughes, L., Holers, V., Deane, K. D., Moreland, L., Filer, A., Pitzalis, C., Forbess, L., Ben-Artzi, A., Salomon-Escoto, K., Raychaudhuri, S., Brenner, M., McDavid, A., Anolik, J., Rao, D., Accelerating Med Partership AMP RA WILEY. 2022: 3442-3444

  View details for Web of Science ID 000877386504234

• Citrullinated Peptide-Specific ACPA Are Present in the Female Genital Tract in Premenopausal Women with and Without RA Marcy, D., Rothfuss, H., Visser, A., Norris, J., Holers, V., Deane, K. D., Robinson, W., Cherrington, B., Demoruelle, K. WILEY. 2022: 1214-1215

  View details for Web of Science ID 000877386501121

• Clonally Expanded Cytotoxic CD8(+) T Cells Recognize Citrullinated Antigens in ACPA(+) Rheumatoid Arthritis Younis, S., Sharpe, O., Rao, N., Carman, J., James, E., Buckner, J., Deane, K. D., Holers, M., Donlin, L., Davis, M., Robinson, W. WILEY. 2022: 3435-3437

  View details for Web of Science ID 000877386504256

• The Association Between Anti-Citrullinated Peptide Antibodies and Subclinical Interstitial Lung Disease in Community-Dwelling Adults: The Multi-Ethnic Study of Atherosclerosis Lanz, J., Hughes-Austin, J., Podolanczuk, A., Kim, J., Austin, J., Robinson, W., Raghu, G., Hoffman, E., Newell, J., Watts, J., Nath, P., Sonavane, S., Barr, R., Bernstein, E. WILEY. 2022: 2364-2366

  View details for Web of Science ID 000877386502215

• An Expanded Anti-citrullinated Protein Antibody Profile Derived Using Unsupervised Machine Learning Predicts Treatment Responses to Biologic Therapies in Rheumatoid Arthritis Aripova, N., Reed, G., England, B., Robinson, W., Pappas, D., Kremer, J., Thiele, G., Mikuls, T. WILEY. 2022: 3185-3186

  View details for Web of Science ID 000877386504105

• An Arthritogenic Strain of Subdoligranulum Specifically Detectable in the Feces of Individuals At-risk for and with Rheumatoid Arthritis Is Bound by ACPA and Stimulates Th17 Cell Activation in Those with Rheumatoid Arthritis Chriswell, M., Rims, C., Maerz, M., Hsu, A., Seifert, J., Feser, M., Bloom, M., Bemis, E., Demoruelle, K., Deane, K. D., Robinson, W., James, E., Buckner, J., Holers, V., Kuhn, K. WILEY. 2022: 1200-1202

  View details for Web of Science ID 000877386501114

• Mechanism-driven strategies for prevention of rheumatoid arthritis. Rheumatology & autoimmunity Holers, V. M., Kuhn, K. A., Demoruelle, M. K., Norris, J. M., Firestein, G. S., James, E. A., Robinson, W. H., Buckner, J. H., Deane, K. D. 2022; 2 (3): 109-119

  Abstract

  In seropositive rheumatoid arthritis (RA), the onset of clinically apparent inflammatory arthritis (IA) is typically preceded by a prolonged period of autoimmunity manifest by the presence of circulating autoantibodies that can include antibodies to citrullinated protein antigens (ACPA) and rheumatoid factor (RF). This period prior to clinical IA can be designated preclinical RA in those individuals who have progressed to a clinical diagnosis of RA, and an 'at-risk' status in those who have not developed IA but exhibit predictive biomarkers of future clinical RA. With the goal of developing RA prevention strategies, studies have characterized immune phenotypes of preclinical RA/at-risk states. From these studies, a model has emerged wherein mucosal inflammation and dysbiosis may lead first to local autoantibody production that should normally be transient, but instead is followed by systemic spread of the autoimmunity as manifest by serum autoantibody elevations, and ultimately drives the development of clinically identified joint inflammation. This model can be envisioned as the progression of disease development through serial 'checkpoints' that in principle should constrain or resolve autoimmunity; however, instead the checkpoints 'fail' and clinical RA develops. Herein we review the immune processes that are likely to be present at each step and the potential therapeutic strategies that could be envisioned to delay, diminish, halt or even reverse the progression to clinical RA. Notably, these prevention strategies could utilize existing therapies approved for clinical RA, therapies approved for other diseases that target relevant pathways in the preclinical/at-risk state, or approaches that target novel pathways.

  View details for DOI 10.1002/rai2.12043

  View details for PubMedID 36312783

• Rheumatoid Arthritis Patient-derived Anti-citrullinated Protein Antibodies (ACPAs) Ameliorate Joint Inflammation in Early Collagen-antibody Induced Arthritis (CAIA) Gomez, A., Brewer, C., Moon, J., Acharya, S., Lanz, T. V., Wang, Q., Min-Oo, G., Niedziela-Majka, A., Robinson, W. WILEY. 2022: 69-70

  View details for Web of Science ID 000877386500045

• Reduction in Rheumatoid Arthritis-Associated Interstitial Lung Disease Risk in Patients Treated with Tofacitinib Baker, M., Liu, Y., Lu, R., Lin, J., Melehani, J., Robinson, W. WILEY. 2022: 4476-4478

  View details for Web of Science ID 000877386506227

• Single-cell Profiling of B Cell Repertoire and Gene Expression in the RA Synovium Reveals Tissue Specific Clonal Expansion Wagner, A., Meednu, N., Dunlap, G., Zhang, F., Jonsson, A., Wei, K., Ben-Artzi, A., Forbess, L., Geraldino-Pardilla, L., Horowitz, D., Hughes, L., Mandelin, A., Salomon-Escoto, K., Tabechian, D., DiCarlo, E., Gravallese, E., Boyce, B., Ritchlin, C., Lederer, J., McGeachy, M., Gregersen, P., Utz, P., Robinson, W., Maecker, H., James, J., Guthridge, J., Perlman, H., Bathon, J., Goodman, S., Firestein, G. S., Boyle, D., Bridges, S., Deane, K. D., Holers, V., Moreland, L., Filer, A., Pitzalis, C., Bykerk, V., Donlin, L., Raychaudhuri, S., Brenner, M., Lupus, A. A., Rao, D., McDavid, A., Anolik, J. WILEY. 2022: 3239-3240

  View details for Web of Science ID 000877386504129

• Mast Cells May Explain Sex Differences in Osteoarthritis Pain Mehta, B., DiCarlo, E., Wang, F., Otero, M., Song, E., Sculco, P., Figgie, M., Goodman, S., Robinson, W., Orange, D. WILEY. 2022: 3265-3267

  View details for Web of Science ID 000877386504140

• CCP plus Immune Checkpoint Inhibitor Arthritis Patients Have Less ACPA Epitope Expansion Than CCP plus Rheumatoid Arthritis Patients Ghosh, N., Rajesh, D., Kirschmann, J., Jannat-Khah, D., Chan, K., Goodman, S., Bykerk, V., Robinson, W., Bass, A. WILEY. 2022: 1528-1530

  View details for Web of Science ID 000877386501282

• SARS-CoV-2 infection of monocytes: balancing acts of antibodies and inflammasomes. Signal transduction and targeted therapy Brewer, R. C., Robinson, W. H., Lanz, T. V. 2022; 7 (1): 250

  View details for DOI 10.1038/s41392-022-01112-w

  View details for PubMedID 35871170

• Mobilization of innate and adaptive antitumor immune responses by the RNP-targeting antibody ATRC-101. Proceedings of the National Academy of Sciences of the United States of America Scholz, A., DeFalco, J., Leung, Y., Aydin, I. T., Czupalla, C. J., Cao, W., Santos, D., Vad, N., Lippow, S. M., Baia, G., Harbell, M., Sapugay, J., Zhang, D., Wu, D., Wechsler, E., Ye, A. Z., Wu, J. W., Peng, X., Vivian, J., Kaplan, H., Collins, R., Nguyen, N., Whidden, M., Kim, D., Millward, C., Benjamin, J., Greenberg, N. M., Serafini, T. A., Emerling, D. E., Steinman, L., Robinson, W. H., Manning-Bog, A. 2022; 119 (19): e2123483119

  Abstract

  SignificanceA target-agnostic approach that harnesses the human antitumor immune response to find potential anticancer lead antibodies and their targets was used to generate ATRC-101, an engineered version of a tumor-targeting antibody identified from a patient with non-small cell lung cancer experiencing an ongoing antitumor immune response. ATRC-101 is an antibody that targets an extracellular, tumor-specific ribonucleoprotein complex. Here, we describe the extracellular binding of this complex and antitumor activity of ATRC-101 in murine models. Preclinical data suggest a mechanism of action in which ATRC-101 activates myeloid cells of the innate immune system, leading to an adaptive immune response that yields its antitumor activity. These data have led to an ongoing phase 1 trial in patients with advanced solid tumors.

  View details for DOI 10.1073/pnas.2123483119

  View details for PubMedID 35507878

• The importance of specific citrullinated clusterin and vimentin found in a multi-coloured bead-based citrulline-peptide array system in rheumatoid arthritis CLINICAL AND EXPERIMENTAL RHEUMATOLOGY Yoshizawa, Y., Nakao, T., Tsuritani, K., Yamada, T., Watanabe, N., Chiba, A., Robinson, W. H., Miyake, S. 2022; 40 (5): 936-944

  View details for Web of Science ID 000805841000010

• Limited Neutralization of Omicron by Antibodies from the BNT162b2 Vaccination against SARS-CoV-2. Research square Lanz, T. V., Brewer, R. C., Jahanbani, S., Robinson, W. H. 2022

  Abstract

  Since early December 2021, the omicron variant has posed additional challenges to the world-wide management of the SARS-CoV-2 pandemic. Immune evasion is a key factor for its increased transmissibility. While serological studies have measured levels of neutralizing antibodies in response to vaccines, our understanding of the humoral immune response to omicron on a single-antibody level is limited. Here, we characterize a set of BNT162b2 vaccine-derived antibodies for neutralization of omicron pseudovirus. We show that approximately 50% of neutralizing anti-RBD antibodies cross-neutralize omicron, albeit with lower potency than the original Wuhan-Hu1 strain. All investigated neutralizing anti-S2 antibodies cross-neutralize omicron, however all of them are less potent than anti-RBD antibodies. While additional booster immunizations of the current vaccine generate increased antibody levels and better protection, we anticipate that the second generation of vaccines will yield more high-affinity antibodies against omicron.

  View details for DOI 10.21203/rs.3.rs-1518378/v1

  View details for PubMedID 35441169

  View details for PubMedCentralID PMC9016652

• KIR+CD8+ T cells suppress pathogenic T cells and are active in autoimmune diseases and COVID-19. Science (New York, N.Y.) Li, J., Zaslavsky, M., Su, Y., Guo, J., Sikora, M. J., van Unen, V., Christophersen, A., Chiou, S., Chen, L., Li, J., Ji, X., Wilhelmy, J., McSween, A. M., Palanski, B. A., Mallajosyula, V. V., Bracey, N. A., Dhondalay, G. K., Bhamidipati, K., Pai, J., Kipp, L. B., Dunn, J. E., Hauser, S. L., Oksenberg, J. R., Satpathy, A. T., Robinson, W. H., Dekker, C. L., Steinmetz, L. M., Khosla, C., Utz, P. J., Sollid, L. M., Chien, Y., Heath, J. R., Fernandez-Becker, N. Q., Nadeau, K. C., Saligrama, N., Davis, M. M. 2022: eabi9591

  Abstract

  Here we find that CD8+ T cells expressing inhibitory killer cell immunoglobulin-like receptors (KIRs) are the human equivalent of Ly49+CD8+ regulatory T cells in mice and are increased in the blood and inflamed tissues of patients with a variety of autoimmune diseases. Moreover, these CD8+ T cells efficiently eliminated pathogenic gliadin-specific CD4+ T cells from celiac disease patients' leukocytes in vitro. We also find elevated levels of KIR+CD8+ T cells, but not CD4+ regulatory T cells, in COVID-19 patients, which correlated with disease severity and vasculitis. Selective ablation of Ly49+CD8+ T cells in virus-infected mice led to autoimmunity post infection. Our results indicate that in both species, these regulatory CD8+ T cells act uniquely to suppress pathogenic T cells in autoimmune and infectious diseases.

  View details for DOI 10.1126/science.abi9591

  View details for PubMedID 35258337

• Antibody cross-reactivity between casein and myelin-associated glycoprotein results in central nervous system demyelination. Proceedings of the National Academy of Sciences of the United States of America Chunder, R., Weier, A., Maurer, H., Luber, N., Enders, M., Luber, G., Heider, T., Spitzer, A., Tacke, S., Becker-Gotot, J., Kurts, C., Iyer, R., Ho, P. P., Robinson, W. H., Lanz, T. V., Kuerten, S. 2022; 119 (10): e2117034119

  Abstract

  Significance Multiple sclerosis (MS) is the most prevalent autoimmune disease of the central nervous system (CNS), leading to irreversible deficits in young adults. Its pathophysiology is believed to be influenced by environmental determinants. As far back as the 1990s, it had been suggested that there is a correlation between the consumption of cow's milk and the prevalence of MS. Here, we not only demonstrate that a high percentage of MS patients harbor antibodies to bovine casein but also that antibody cross-reactivity between cow's milk and CNS antigens can exacerbate demyelination. Our data broaden the current understanding of how diet influences the etiology of MS and set the stage for combining personalized diet plans with disease-modifying treatment strategies.

  View details for DOI 10.1073/pnas.2117034119

  View details for PubMedID 35235454

• Extracellular histones aggravate autoimmune arthritis by lytic cell death. Frontiers in immunology Jung, J., Lee, L. E., Kim, H., Kim, J. E., Jang, S. H., Roh, J. S., Lee, B., Robinson, W. H., Sohn, D. H., Pyun, J., Song, J. J. 2022; 13: 961197

  Abstract

  Although recent studies have demonstrated a proinflammatory effect of extracellular histones in sepsis via endothelial cytotoxicity, little is known about their contribution to autoimmune arthritis. Therefore, we investigated the role of extracellular histones in autoimmune arthritis and their cytotoxic effect on synoviocytes and macrophages. We measured histones in the synovial fluid of patients with rheumatoid arthritis (RA) and evaluated arthritis severity in a serum-transfer arthritis (STA) mouse model with intraperitoneal histone injection. Histone-induced cytotoxicity was measured using SYTOX green staining in the synoviocyte cell line MH7A and macrophages differentiated from the monocytic cell line THP-1, and the production of damage-associated molecular patterns (DAMPs) was measured by HMGB1 and ATP. Furthermore, we performed RNA-seq analysis of THP-1 cells stimulated with H2B-alpha1 peptide or with its citrullinated form. The levels of histones were elevated in RA synovial fluid, and histones aggravated arthritis in the STA model. Histones induced cytotoxicity and DAMP production in synoviocytes and macrophages. Chondroitin sulfate reduced histone-induced cytotoxicity, while lipopolysaccharides aggravated cytotoxicity. Moreover, the cytotoxicity decreased when the arginines in H2B-alpha1 were replaced with citrullines, which demonstrated its electrostatic nature. In transcriptome analysis, H2B-alpha1 changed the gene expression pattern of THP-1 cells involving chemokines, interleukin-1, -4, -10, -13, and toll-like receptor (TLR) signaling pathways. Extracellular histones were increased in RA synovial fluid and aggravated synovitis in STA. They induced lytic cell death through electrostatic interaction with synoviocytes and macrophages, leading to the secretion of DAMPs. These findings suggest that histones play a central role in autoimmune arthritis.

  View details for DOI 10.3389/fimmu.2022.961197

  View details for PubMedID 36032105

• Differential Changes in ACPA Fine Specificity and Gene Expression in a Randomized Trial of Abatacept and Adalimumab in Rheumatoid Arthritis. Rheumatology and therapy Jabado, O., Maldonado, M. A., Schiff, M., Weinblatt, M. E., Fleischmann, R., Robinson, W. H., He, A., Patel, V., Greenfield, A., Saini, J., Galbraith, D., Connolly, S. E. 2021

  Abstract

  INTRODUCTION: The biologics abatacept and adalimumab have different mechanisms of action (MoAs). We analyzed data from patients with rheumatoid arthritis treated in AMPLE (NCT00929864) to explore the pharmacodynamic effects of abatacept or adalimumab on anti-citrullinated protein antibodies (ACPAs) and gene expression.METHODS: AMPLE was a phase IIIb, 2-year, randomized, head-to-head trial of abatacept versus adalimumab. Post hoc analyses of baseline anti-cyclic citrullinated peptide-2 (anti-CCP2, an ACPA surrogate) positive (+) status and ACPA fine-specificity profiles over time, as well as transcriptional profiling (peripheral whole blood), were performed.RESULTS: Of 646 patients treated (abatacept, n=318; adalimumab, n=328), ACPA and gene expression data were available from 508 and 566 patients, respectively. In anti-CCP2+patients (n=388), baseline fine specificities for most ACPAs were highly correlated; over 2years, levels decreased with abatacept but not adalimumab. By year 2, expression of genes associated with T cell co-stimulation and antibody production was lower for abatacept versus adalimumab; expression of genes associated with proinflammatory signaling was lower for adalimumab versus abatacept. Treatment modulated the expression of T- and B-cell gene signatures, with differences in CD8+T cells, activated T cells, plasma cells, B cells, natural killer cells (all lower with abatacept versus adalimumab), and polymorphonuclear leukocytes (higher with abatacept versus adalimumab).CONCLUSIONS: In AMPLE, despite similar clinical outcomes, data showed that pharmacodynamic/genetic changes after 2years of abatacept or adalimumab were consistent with drug MoAs. Further assessment of the relationship between such changes and clinical outcomes, including prediction of response, is warranted.TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT00929864.

  View details for DOI 10.1007/s40744-021-00404-x

  View details for PubMedID 34878629

• Histologic subtype of cutaneous immune-related adverse events predicts overall survival in patients receiving immune checkpoint inhibitors. Journal of the American Academy of Dermatology Hirotsu, K. E., Scott, M. K., Marquez, C., Tran, A. T., Rieger, K. E., Novoa, R. A., Robinson, W. H., Kwong, B. Y., Zaba, L. C. 2021

  View details for DOI 10.1016/j.jaad.2021.11.050

  View details for PubMedID 34875301

• Antibody Responses to Epstein-Barr Virus in the Preclinical Period of Rheumatoid Arthritis Suggest the Presence of Increased Viral Reactivation Cycles. Arthritis & rheumatology (Hoboken, N.J.) Fechtner, S., Berens, H., Bemis, E., Johnson, R. L., Guthridge, C. J., Carlson, N. E., Demoruelle, M. K., Harley, J. B., Edison, J. D., Norris, J. A., Robinson, W. H., Deane, K. D., James, J. A., Holers, V. M. 2021

  Abstract

  OBJECTIVE: Patients with established rheumatoid arthritis (RA) demonstrate altered immune responses to Epstein-Barr virus (EBV), but the presence and role(s) of EBV have not been fully explored in the preclinical period. We hypothesized that EBV infection, as evidenced by an altered anti-EBV antibody response, could either play an important role in driving the development or be a result of expanded RA-related autoimmunity.METHODS: 83 subjects with RA based on 1987 ACR criteria and 83 matched controls were evaluated. Subject and matched control sera from the pre and post- RA diagnosis periods were tested for 5 anti-EBV antibodies (EBNA-1-IgG, VCA-IgG, EA-IgG, VCA-IgA, and EA-IgA), 7 RA-related autoantibodies (RF-neph, RF-IgA, RF-IgM, RF-IgG, CCP2, CCP3, CCP3.1), 22 cytokine/chemokine, 36 individual APCAs, and CMV-IgG antibodies. Random forest classification, mixed and joint mixed modelling were used to determine differences in anti-EBV antibodies between RA cases and controls.RESULTS: Random Forest analysis identified preclinical EBV antibodies that differentiate RA subjects from controls. Specifically, EA-IgG antibody levels are higher in RA cases (0.82 ± 0.72) compared to controls (0.49 ± 0.28). Elevations in EA-IgG levels significantly correlated with increasing RF-IgM levels in future RA cases (p = 0.007) but not in controls (p = 0.150). CMV-IgG antibody levels did not differ between groups.CONCLUSION: Subjects who eventually develop classified RA demonstrate elevated EA-IgG antibody levels in the preclinical period, which suggests the presence of increased EBV re-activation cycles. A combination of RF and EBV reactivation may play an important role in the development of RA.

  View details for DOI 10.1002/art.41994

  View details for PubMedID 34605217

• New-onset IgG autoantibodies in hospitalized patients with COVID-19. Nature communications Chang, S. E., Feng, A., Meng, W., Apostolidis, S. A., Mack, E., Artandi, M., Barman, L., Bennett, K., Chakraborty, S., Chang, I., Cheung, P., Chinthrajah, S., Dhingra, S., Do, E., Finck, A., Gaano, A., GeSSner, R., Giannini, H. M., Gonzalez, J., Greib, S., Gundisch, M., Hsu, A. R., Kuo, A., Manohar, M., Mao, R., Neeli, I., Neubauer, A., Oniyide, O., Powell, A. E., Puri, R., Renz, H., Schapiro, J., Weidenbacher, P. A., Wittman, R., Ahuja, N., Chung, H., Jagannathan, P., James, J. A., Kim, P. S., Meyer, N. J., Nadeau, K. C., Radic, M., Robinson, W. H., Singh, U., Wang, T. T., Wherry, E. J., Skevaki, C., Luning Prak, E. T., Utz, P. J. 2021; 12 (1): 5417

  Abstract

  COVID-19 is associated with a wide range of clinical manifestations, including autoimmune features and autoantibody production. Here we develop three protein arrays to measure IgG autoantibodies associated with connective tissue diseases, anti-cytokine antibodies, and anti-viral antibody responses in serum from 147 hospitalized COVID-19 patients. Autoantibodies are identified in approximately 50% of patients but in less than 15% of healthy controls. When present, autoantibodies largely target autoantigens associated with rare disorders such as myositis, systemic sclerosis and overlap syndromes. A subset of autoantibodies targeting traditional autoantigens or cytokines develop de novo following SARS-CoV-2 infection. Autoantibodies track with longitudinal development of IgG antibodies recognizing SARS-CoV-2 structural proteins and a subset of non-structural proteins, but not proteins from influenza, seasonal coronaviruses or other pathogenic viruses. We conclude that SARS-CoV-2 causes development of new-onset IgG autoantibodies in a significant proportion of hospitalized COVID-19 patients and are positively correlated with immune responses to SARS-CoV-2 proteins.

  View details for DOI 10.1038/s41467-021-25509-3

  View details for PubMedID 34521836

• Effective Viral Vector SARS-CoV-2 Booster Vaccination in a Patient with Rheumatoid Arthritis after Initial Ineffective mRNA Vaccine Response. Arthritis & rheumatology (Hoboken, N.J.) Baker, M. C., Mallajosyula, V., Davis, M. M., Boyd, S. D., Nadeau, K. C., Robinson, W. H. 2021

  Abstract

  Managing patients with rheumatic disease during the COVID-19 pandemic has posed a unique challenge. Immunosuppressed patients are at an increased risk for developing severe COVID-19 and may not derive full protection from the vaccine (1-5). Thus, it is paramount we develop strategies whereby rheumatic disease patients can be protected from the pandemic virus and its variants.

  View details for DOI 10.1002/art.41978

  View details for PubMedID 34514750

• Symptom Trajectories in the Transition from Pre-Rheumatoid Arthritis to Clinically-Apparent Inflammatory Arthritis Haville, S., Seifert, J., Barzideh, S., Moss, L., Rao, N., Johnsen, A., Buckner, J., James, E., Posso, S., Firestein, G., Boyle, D., Robinson, W., Holers, V., Deane, K. WILEY. 2021: 3444-3445

  View details for Web of Science ID 000744545206217

• Identifying Trajectories and Endotypes in the Evolution of Pre-Rheumatoid Arthritis with Autoantibody Testing and Artificial Adaptive System Analysis Buscema, P., Massini, G., Della Torre, F., Asadi-Zeydabadi, M., O'Donnell, C., Newman, F., Tagg, R., Lodwick, W., Collora, C., Feser, M., Moss, L., Robinson, W., Thiele, G., Mikuls, T., Edison, J., Holers, V., Deane, K. WILEY. 2021: 3442-3444

  View details for Web of Science ID 000744545206216

• Plasmablast-derived Autoantibodies from Individuals At-risk for RA That Target RA-relevant Antigens Are Polyreactive with Arthritogenic Bacteria Chriswell, M., Seifert, J., Bloom, M., Rims, C., Feser, M., Deane, K., Norris, J., James, E., Buckner, J., Robinson, W., Holers, V., Kuhn, K. WILEY. 2021: 953-954

  View details for Web of Science ID 000744545201216

• Osteoarthritis Risk Is Increased in Patients with Atopic Disease Baker, M., Sheth, K., Lu, R., Lu, D., von Kaeppler, E., Bhat, A., Felson, D., Robinson, W. WILEY. 2021: 2351-2353

  View details for Web of Science ID 000744545204174

• New-Onset IgG Autoantibodies in Hospitalized Patients with COVID-19 Chang, S., Feng, A., Meng, W., Apostolidis, S., Mack, E., Artandi, M., Barman, L., Bennett, K., Chakraborty, S., Chang, I., Cheung, P., Chinthrajah, S., Dhingra, S., Do, E., Finck, A., Gaano, A., Gessner, R., Giannini, H., Gonzalez, J., Greib, S., Gundisch, M., Hsu, A., Kuo, A., Manohar, M., Mao, R., Neeli, I., Neubauer, A., Oniyide, O., Powell, A., Puri, R., Renz, H., Schapiro, J., Weidenbacher, P., Wittman, R., Ahuja, N., Chung, H., Jagannathan, P., James, J., Kim, P., Meyer, N., Nadeau, K., Radic, M., Robinson, W., Singh, U., Wang, T., Wherry, J., Skevaki, C., Prak, E., Utz, P. WILEY. 2021: 3202-3205

  View details for Web of Science ID 000744545206097

• Unsupervised Clustering of Histology and Ultrasound Scores Identifies Osteoarthritis Subtypes Konnaris, M., Bell, R., Pannellini, T., DiCarlo, E., Gibbons, J., Nwawka, O., Lee, S., Sculco, P., Parks, M., Figgie, M., Donlin, L., Orange, D., Sculco, T., Robinson, W., Goodman, S., Otero, M., Mehta, B. WILEY. 2021: 421-424

  View details for Web of Science ID 000744545200213

• Single-cell Profiling of B and T Cell Repertoire and Gene Expression in the RA Synovium Reveals Tissue Specific Clonal Expansion Meednu, N., Wagner, A., Dunlap, G., Zhang, F., Jonsson, A., Wei, K., Utz, P., Robinson, W., Maecker, H., James, J., Guthridge, J., Bridges, S., Bykerk, V., Donlin, L., Goodman, S., DiCarlo, E., Ritchlin, C., Tabechian, D., Lederer, J., Gravallese, E., McGeachy, M., Firestein, G., Boyle, D., Gregersen, P., Horowitz, D., Perlman, H., Mandelin, A., Bathon, J., Geraldino--Pardilla, L., Hughes, L., Holers, V., Deane, K., Moreland, L., Filer, A., Pitzalis, C., Forbess, L., Ben-Artzi, A., Salomon-Escoto, K., Raychaudhuri, S., Brenner, M., Rao, D., McDavid, A., Anolik, J., Medicines, A. WILEY. 2021: 951-952

  View details for Web of Science ID 000744545201215

• Autoantibodies and the Risk of Incident Cardiovascular Disease in US Veterans with Rheumatoid Arthritis Johnson, T., Duryee, M., Hunter, C., Roul, P., Yang, Y., Sokolove, J., Robinson, W., Baker, J., Thiele, G., Mikuls, T., England, B. WILEY. 2021: 547-550

  View details for Web of Science ID 000744545201024

• Metformin Use Reduces the Risk of Developing Osteoarthritis: A Propensity Score Matching Study Sheth, K., Lu, D., Lu, R., Robinson, W., Baker, M. WILEY. 2021: 994-996

  View details for Web of Science ID 000744545201237

• Tendon-Derived Progenitor Cells With Multilineage Potential Are Present Within Human Patellar Tendon. Orthopaedic journal of sports medicine Leonardi, E. A., Xiao, M., Murray, I. R., Robinson, W. H., Abrams, G. D. 2021; 9 (8): 23259671211023452

  Abstract

  Background: Progenitor cells serve as a promising source of regenerative potential in a variety of tissue types yet remain underutilized in tendinopathy. Tendon-derived progenitor cells (TDPCs) have previously been isolated from hamstring tendon but only as part of a concomitant medical procedure. Determining the presence of TDPCs in patellar tendon may facilitate clinical utilization of these cells because of the relative accessibility of this location for tissue harvest.Purpose: To characterize TDPCs in human patellar tendon samples.Study Design: Descriptive laboratory study.Methods: Human patellar tendon samples were obtained during elective knee surgery. TDPCs were isolated and seeded at an optimal low cell density and subcultured to confluence for up to 2 passages. Flow cytometry was used to analyze for the expression of CD90+, CD105+, CD44+, and CD31-, CD34-, and CD45- markers. The multilineage differentiation potential of TDPCs was tested in vitro via adipogenic, osteogenic, and chondrogenic culture with subsequent cytochemical staining for Oil Red O, Alizarin Red, and Alcian Blue, respectively. Enzyme-linked immunosorbent assay was used to quantify the amount of adiponectin, alkaline phosphatase, and SRY-box transcription factor 9 secreted into cell culture supernatant for further confirmation of lineage differentiation. Results were analyzed statistically using the 2-tailed Student t test.Results: TDPCs demonstrated near-uniform expression of CD90, CD105, and CD44 with minimal expression of CD34, CD31, and CD45. Adipogenic, osteogenic, and chondrogenic differentiation of TDPCs was confirmed using qualitative analysis. The expression of adiponectin, alkaline phosphatase, and SRY-box transcription factor 9 were significantly increased in differentiated cells versus undifferentiated TDPCs (P < .05).Conclusion: TDPCs can be successfully isolated from human patellar tendon samples, and they exhibit characteristics of multipotent progenitor cells.Clinical Relevance: These data demonstrate the promise of patellar tendon tissue as a source of progenitor cells for use in biologic therapies for the treatment of tendinopathy.

  View details for DOI 10.1177/23259671211023452

  View details for PubMedID 34435068

• The importance of specific citrullinated clusterin and vimentin found in a multi-coloured bead-based citrulline-peptide array system in rheumatoid arthritis. Clinical and experimental rheumatology Yoshizawa, Y., Nakao, T., Tsuritani, K., Yamada, T., Watanabe, N., Chiba, A., Robinson, W. H., Miyake, S. 2021

  Abstract

  OBJECTIVES: The importance of citrullination in rheumatoid arthritis (RA) has been reported, but the degree to which individual citrullinated proteins affect the onset and progression of RA is still unclear. We aimed to identify citrullinated proteins that may play an important role in the onset and progression of RA using an individualised anti-citrullinated protein antibody (ACPA) evaluation system with citrullinated peptides as probes.METHODS: Serum samples from 50 normal donors and 51 RA patients were evaluated using a custom MagPlexTM bead array with 13 types of citrullinated peptide. The presence/absence of ACPAs that react to each citrullinated peptide in each subject was determined using the Z-score, which was calculated based on the fluorescence intensity distribution of a sample from a normal donor. Whether the fluorescence intensity was inhibited when free citrullinated peptides were added to a system was also evaluated.RESULTS: Median fluorescence intensities obtained from beads coupled with the 13 types of citrullinated peptide were all significantly higher in RA patients versus normal donors. With a Z-score ≥2 as the cut-off value for the presence of ACPAs, ACPAs that recognised five types of citrullinated peptides derived from fibrinogen A, fillagrin, clusterin, and vimentin were widely detected in RA patients. In addition, inhibition experiments showed that citrullinated vimentin, clusterin, and enolase 1A peptides inhibited coupling of ACPAs to other citrullinated peptides.CONCLUSIONS: ACPAs to many citrullinated proteins exhibited cross-reactivity to citrullinated clusterin and vimentin, suggesting the importance of citrullinated clusterin and vimentin in the early stages of RA pathogenesis.

  View details for PubMedID 34251306

• First-in-human phase 1b study of ATRC-101, a patient-derived antibody with a tumor-specific target, as monotherapy or in combination with pembrolizumab, in patients with solid tumors. Benjamin, J. E., Higgins, N., Millward, C., DeFalco, J., Manning-Bog, A., Aydin, I. T., Zhang, D., Lippow, S. M., Scholz, A., Leung, Y., Zhu, Y., Sedello, A., Rogers, Z., Dhawan, I. K., Emerling, D., Robinson, W. H., Serafini, T. A., Greenberg, N. M. AMER ASSOC CANCER RESEARCH. 2021

  View details for Web of Science ID 000680263501286

• Pathogenic Role of Circulating Citrullinated Antigens and Anti-Cyclic Monoclonal Citrullinated Peptide Antibodies in Rheumatoid Arthritis FRONTIERS IN IMMUNOLOGY Won, P., Kim, Y., Jung, H., Rim, Y., Sohn, D., Robinson, W. H., Moon, S., Ju, J. 2021; 12: 692242

  Abstract

  We examined whether it is possible to directly detect citrullinated antigens in the serum of rheumatoid arthritis (RA) patients using a monoclonal antibody (mAb) designed to be specific for citrullinated peptides. In order to confirm the potential of the mAb as a direct arthritis-inducing substance through experimental model of RA, a monoclonal antibody (mAb) 12G1 was generated using by immunization of mice with a challenging cyclic citrullinated peptide. Immunohistochemical analysis of RA-affected synovial tissue showed that our mAb 12G1 could indeed detect citrullinated proteins in target tissues. Subsequently, serum levels of citrullinated type II collagen and filaggrin were measured in healthy volunteers, patients with RA, ankylosing spondylitis (AS), and systemic lupus erythematosus (SLE) using a 12G1-based sandwich ELISA. This showed that citrullinated filaggrin showed 78.9% sensitivity and 85.9% specificity for RA diagnosis with a cutoff optical density (OD) value of 1.013, comparable with the results from a second-generation anti-citrullinated protein antibody (ACPA) test. Circulating citrullinated collagen and filaggrin were detected even in sera of RA patients who were negative for both rheumatoid factor (RF) and ACPA. ELISA results also showed that RF and ACPA titers showed significantly positive correlation with both citrullinated collagen and filaggrin OD values in sera of RA patients. 12G1 challenging aggravated the severity of murine arthritis. In summary, mAb 12G1 can directly detect citrullinated proteins in RA target tissue and in sera of RA patients and 12G1 showed direct arthritogenic potential in vivo. This, 12G1 might be useful for diagnosis of RA including seronegative RA and may help to elucidate the pathophysiological role of citrullination in RA.

  View details for DOI 10.3389/fimmu.2021.692242

  View details for Web of Science ID 000675002500001

  View details for PubMedID 34305925

  View details for PubMedCentralID PMC8294326

• Interleukin 4 promotes anti-inflammatory macrophages that clear cartilage debris and inhibits osteoclast development to protect against osteoarthritis. Clinical immunology (Orlando, Fla.) von Kaeppler, E. P., Wang, Q., Raghu, H., Bloom, M. S., Wong, H., Robinson, W. H. 2021: 108784

  Abstract

  OBJECTIVE: Osteoarthritis (OA), the leading cause of joint failure, is characterized by breakdown of articular cartilage and remodeling of subchondral bone in synovial joints. Despite the high prevalence and debilitating effects of OA, no disease-modifying drugs exist. Increasing evidence, including genetic variants of the interleukin 4 (IL-4) and IL-4 receptor genes, implicates a role for IL-4 in OA, however, the mechanism underlying IL-4 function in OA remains unknown. Here, we investigated the role of IL-4 in OA pathogenesis.METHODS: Il4-, myeloid-specific-Il4ra-, and Stat6-deficient and control mice were subjected to destabilization of the medial meniscus to induce OA. Macrophages, osteoclasts, and synovial explants were stimulated with IL-4 in vitro, and their function and expression profiles characterized.RESULTS: Mice lacking IL-4, IL-4Ra in myeloid cells, or STAT6 developed exacerbated cartilage damage and osteophyte formation relative to WT controls. In vitro analyses revealed that IL-4 downregulates osteoarthritis-associated genes, enhances macrophage phagocytosis of cartilage debris, and inhibits osteoclast differentiation and activation via the type I receptor.CONCLUSION: Our findings demonstrate that IL-4 protects against osteoarthritis in a myeloid and STAT6-dependent manner. Further, IL-4 can promote an immunomodulatory microenvironment in which joint-resident macrophages polarize towards an M2 phenotype and efficiently clear pro-inflammatory debris, and osteoclasts maintain a homeostatic level of activity in subchondral bone. These findings support a role for IL-4 modulation of myeloid cell types in maintenance of joint health and identify a pathway that could provide therapeutic benefit for osteoarthritis.

  View details for DOI 10.1016/j.clim.2021.108784

  View details for PubMedID 34126239

• Dual IgA/IgG family plasmablast-derived mAbs from peripheral blood of individuals at risk for rheumatoid arthritis are polyreactive to autoantigens and intestinal bacteria Chriswell, M., Bloom, M., Seifert, J. A., Feser, M., Demoruelle, M., Deane, K. D., Norris, J. M., Robinson, W. H., Holers, V., Kuhn, K. A. AMER ASSOC IMMUNOLOGISTS. 2021

  View details for Web of Science ID 000713665800407

• Associations of serum cytokines and chemokines with the risk of incident cancer in a prospective rheumatoid arthritis cohort. International immunopharmacology England, B. R., Campany, M., Sayles, H., Roul, P., Yang, Y., Ganti, A. K., Sokolove, J., Robinson, W. H., Reimold, A. M., Kerr, G. S., Cannon, G. W., Sauer, B. C., Baker, J. F., Thiele, G. M., Mikuls, T. R. 2021; 97: 107719

  Abstract

  OBJECTIVES: We aimed to assess whether serum cytokine/chemokine concentrations predict incident cancer in RA patients.METHODS: Data from cancer-free enrollees in the Veterans Affairs Rheumatoid Arthritis (VARA) Registry were linked to a national VA oncology database and the National Death Index (NDI) to identify incident cancers. Seventeen serum cytokines/chemokines were measured from enrollment serum and an overall weighted cytokine/chemokine score (CK score) was calculated. Associations of cytokines/chemokines with all-site, lung, and lymphoproliferative cancers were assessed in Cox regression models accounting for relevant covariates including age, sex, RA disease activity, and smoking.RESULTS: In 1216 patients, 146 incident cancers (42 lung and 23 lymphoproliferative cancers) occurred over 10,072 patient-years of follow-up with a median time of 4.6years from enrollment (cytokine/chemokine measurement) to cancer incidence. In fully adjusted models, CK score was associated with a higher risk of all-site (aHR 1.32, 95% CI 1.01-1.71, p<0.001), lung (aHR 1.81, 1.40-2.34, p=0.001), and lung/lymphoproliferative (aHR 1.54 [1.35-1.75], p<0.001) cancer. The highest quartile of CK score was associated with a higher risk of all-site (aHR 1.91, 0.96-3.81, p=0.07; p-trend=0.005), lung (aHR 8.18, 1.63-41.23, p=0.01; p-trend<0.001), and lung/lymphoproliferative (aHR 4.56 [1.84-11.31], p=0.001; p-trend<0.001) cancer. Thirteen of 17 individual analytes were associated with incident cancer risk.CONCLUSION: Elevated cytokine/chemokine concentrations are predictive of future cancer in RA patients, particularly lung and lymphoproliferative cancers. These results suggest that the measurement of circulating cytokines/chemokines could be informative in cancer risk stratification and could provide insight into future cancer prevention strategies in RA, and possibly individuals without RA.

  View details for DOI 10.1016/j.intimp.2021.107719

  View details for PubMedID 33933845

• Predicting Lyme Disease From Patients' Peripheral Blood Mononuclear Cells Profiled With RNA-Sequencing FRONTIERS IN IMMUNOLOGY Clarke, D. B., Rebman, A. W., Bailey, A., Wojciechowicz, M. L., Jenkins, S. L., Evangelista, J. E., Danieletto, M., Fan, J., Eshoo, M. W., Mosel, M. R., Robinson, W., Ramadoss, N., Bobe, J., Soloski, M. J., Aucott, J. N., Ma'ayan, A. 2021; 12: 636289

  Abstract

  Although widely prevalent, Lyme disease is still under-diagnosed and misunderstood. Here we followed 73 acute Lyme disease patients and uninfected controls over a period of a year. At each visit, RNA-sequencing was applied to profile patients' peripheral blood mononuclear cells in addition to extensive clinical phenotyping. Based on the projection of the RNA-seq data into lower dimensions, we observe that the cases are separated from controls, and almost all cases never return to cluster with the controls over time. Enrichment analysis of the differentially expressed genes between clusters identifies up-regulation of immune response genes. This observation is also supported by deconvolution analysis to identify the changes in cell type composition due to Lyme disease infection. Importantly, we developed several machine learning classifiers that attempt to perform various Lyme disease classifications. We show that Lyme patients can be distinguished from the controls as well as from COVID-19 patients, but classification was not successful in distinguishing those patients with early Lyme disease cases that would advance to develop post-treatment persistent symptoms.

  View details for DOI 10.3389/fimmu.2021.636289

  View details for Web of Science ID 000631064000001

  View details for PubMedID 33763080

  View details for PubMedCentralID PMC7982722

• Diminished cytokine-induced Jak/STAT signaling is associated with rheumatoid arthritis and disease activity. PloS one Ptacek, J. n., Hawtin, R. E., Sun, D. n., Louie, B. n., Evensen, E. n., Mittleman, B. B., Cesano, A. n., Cavet, G. n., Bingham, C. O., Cofield, S. S., Curtis, J. R., Danila, M. I., Raman, C. n., Furie, R. A., Genovese, M. C., Robinson, W. H., Levesque, M. C., Moreland, L. W., Nigrovic, P. A., Shadick, N. A., O'Dell, J. R., Thiele, G. M., Clair, E. W., Striebich, C. C., Hale, M. B., Khalili, H. n., Batliwalla, F. n., Aranow, C. n., Mackay, M. n., Diamond, B. n., Nolan, G. P., Gregersen, P. K., Bridges, S. L. 2021; 16 (1): e0244187

  Abstract

  Rheumatoid arthritis (RA) is a systemic and incurable autoimmune disease characterized by chronic inflammation in synovial lining of joints. To identify the signaling pathways involved in RA, its disease activity, and treatment response, we adapted a systems immunology approach to simultaneously quantify 42 signaling nodes in 21 immune cell subsets (e.g., IFNα→p-STAT5 in B cells) in peripheral blood mononuclear cells (PBMC) from 194 patients with longstanding RA (including 98 patients before and after treatment), and 41 healthy controls (HC). We found multiple differences between patients with RA compared to HC, predominantly in cytokine-induced Jak/STAT signaling in many immune cell subsets, suggesting pathways that may be associated with susceptibility to RA. We also found that high RA disease activity, compared to low disease activity, was associated with decreased (e.g., IFNα→p-STAT5, IL-10→p-STAT1) or increased (e.g., IL-6→STAT3) response to stimuli in multiple cell subsets. Finally, we compared signaling in patients with established, refractory RA before and six months after initiation of methotrexate (MTX) or TNF inhibitors (TNFi). We noted significant changes from pre-treatment to post-treatment in IFNα→p-STAT5 signaling and IL-10→p-STAT1 signaling in multiple cell subsets; these changes brought the aberrant RA signaling profiles toward those of HC. This large, comprehensive functional signaling pathway study provides novel insights into the pathogenesis of RA and shows the potential of quantification of cytokine-induced signaling as a biomarker of disease activity or treatment response.

  View details for DOI 10.1371/journal.pone.0244187

  View details for PubMedID 33444321

• Serologic response to Borrelia antigens varies with clinical phenotype in children and young adults with Lyme disease. Journal of clinical microbiology Radtke, F. A., Ramadoss, N., Garro, A. C., Bennett, J. E., Levas, M. N., Robinson, W. H., Nigrovic, P. A., Nigrovic, L. E., Net, P. L. 2021: JCM0134421

  Abstract

  Lyme disease is commonly diagnosed by serologic response to Borrelia burgdorferi and related species, but the relationship between serologic targets and clinical features is unknown. We developed a multi-antigen Luminex-based panel and evaluated IgG responses in 527 children 1 to 21 years of age assessed for Lyme disease across 4 Pedi Lyme Net emergency departments, including 127 Lyme cases defined by either an erythema migrans (EM) lesion or positive C6 enzyme immunoassay followed by immunoblot and 400 patients considered clinical mimics. Of 42 antigens tested, 26 elicited specific reactivity in Lyme patients, without marked age-dependent variation. Children with single EM lesions typically lacked Borrelia-specific IgG. By principal component analysis, children with early disseminated and late Lyme disease clustered separately from clinical mimics and also from each other. Neurological disease and arthritis exhibited distinct serologic responses, with OspC variants overrepresented in neurological disease and p100, BmpA, p58 and p45 overrepresented in arthritis. Machine learning identified a 3-antigen panel (VlsE_Bb, p41_Bb, OspC_Bafz) that distinguished Lyme disease from clinical mimics with a sensitivity of 86.6% (95% confidence interval [CI] 80.3-92.1) and a specificity of 95.5% (95% CI 93.4-97.4). Sensitivity was much lower in early Lyme disease (38.5%, 95% CI 15.4-69.2). Interestingly, 17 children classified as Lyme mimics had a positive 3-antigen panel, suggesting that more comprehensive serologic analysis could help refine Lyme diagnosis. In conclusion, multiplex antigen panels provide a novel approach to understanding the immune response in Lyme disease, potentially helping to facilitate accurate diagnosis and to understand differences between clinical stages.

  View details for DOI 10.1128/JCM.01344-21

  View details for PubMedID 34379528

• Molecular Microbiological and Immune Characterization of a Cohort of Patients Diagnosed with Early Lyme Disease JOURNAL OF CLINICAL MICROBIOLOGY Mosel, M. R., Rebman, A. W., Carolan, H. E., Montenegro, T., Lovari, R., Schutzer, S. E., Ecker, D. J., Yang, T., Ramadoss, N. S., Robinson, W. H., Soloski, M. J., Eshoo, M. W., Aucott, J. N. 2021; 59 (1)

  View details for DOI 10.1128/JCM.00515-20

  View details for Web of Science ID 000600837900003

• Modeling human adaptive immune responses with tonsil organoids. Nature medicine Wagar, L. E., Salahudeen, A. n., Constantz, C. M., Wendel, B. S., Lyons, M. M., Mallajosyula, V. n., Jatt, L. P., Adamska, J. Z., Blum, L. K., Gupta, N. n., Jackson, K. J., Yang, F. n., Röltgen, K. n., Roskin, K. M., Blaine, K. M., Meister, K. D., Ahmad, I. N., Cortese, M. n., Dora, E. G., Tucker, S. N., Sperling, A. I., Jain, A. n., Davies, D. H., Felgner, P. L., Hammer, G. B., Kim, P. S., Robinson, W. H., Boyd, S. D., Kuo, C. J., Davis, M. M. 2021

  Abstract

  Most of what we know about adaptive immunity has come from inbred mouse studies, using methods that are often difficult or impossible to confirm in humans. In addition, vaccine responses in mice are often poorly predictive of responses to those same vaccines in humans. Here we use human tonsils, readily available lymphoid organs, to develop a functional organotypic system that recapitulates key germinal center features in vitro, including the production of antigen-specific antibodies, somatic hypermutation and affinity maturation, plasmablast differentiation and class-switch recombination. We use this system to define the essential cellular components necessary to produce an influenza vaccine response. We also show that it can be used to evaluate humoral immune responses to two priming antigens, rabies vaccine and an adenovirus-based severe acute respiratory syndrome coronavirus 2 vaccine, and to assess the effects of different adjuvants. This system should prove useful for studying critical mechanisms underlying adaptive immunity in much greater depth than previously possible and to rapidly test vaccine candidates and adjuvants in an entirely human system.

  View details for DOI 10.1038/s41591-020-01145-0

  View details for PubMedID 33432170

• Associations between an expanded autoantibody profile and treatment responses to biologic therapies in patients with rheumatoid arthritis. International immunopharmacology Petro, A. D., Dougherty, J., England, B. R., Sayles, H., Duryee, M. J., Hunter, C. D., Kremer, J. M., Pappas, D. A., Robinson, W. H., Curtis, J. R., Thiele, G. M., Mikuls, T. R. 2020; 91: 107260

  Abstract

  BACKGROUND: Although biologics represent a major advance in rheumatoid arthritis (RA), many patients fail to achieve adequate responses to these agents. We examined whether combined positivity to three well-characterized autoantibodies predicts treatment response among RA patients initiating biologics.METHODS: The study included biologic-naive patients initiating anti-TNF treatment, biologic-exposed patients switching to rituximab or tocilizumab, and patients (biologic naive or exposed) initiating abatacept. Rheumatoid factor (RF), anti-cyclic citrullinated peptide (CCP) antibody, and IgG antibodies to malondialdehyde-acetaldehyde (MAA) were measured using banked enrollment serum. The relationship between the number of autoantibodies positive (0-3) and treatment response (absolute improvement in 28-joint Disease Activity Score [DAS28-CRP] or improvement>1.2) at 6months was examined using multivariable linear and logistic regression.RESULTS: Of 1,229 patients initiating biologics, 79% were women; 89% were Caucasian. The number of baseline RA-related autoantibodies positive was associated with improved treatment response in a dose-dependent fashion. Compared to patients seronegative for all autoantibodies, adjusting for covariates, those positive for all three were more than twice (OR 2.35; 95% CI 1.57-3.51) as likely to achieve DAS28 improvement>1.2 units. Associations of autoantibody positivity with biologic treatment response were strongest for anti-CCP antibody, persisted in analyses limited to biologic naive patients, and did not appear to differ markedly among different agents examined.CONCLUSION: An expanded autoantibody profile appears to significantly predict RA treatment response to biologic treatment in a dose-dependent fashion. Incorporating these serologic profiles with additional biomarkers or other informative patient characteristics could provide an opportunity to personalize RA management.

  View details for DOI 10.1016/j.intimp.2020.107260

  View details for PubMedID 33360371

• Molecular Microbiological and Immune Characterization of a Cohort of Patients Diagnosed with Early Lyme Disease. Journal of clinical microbiology Mosel, M. R., Rebman, A. W., Carolan, H. E., Montenegro, T., Lovari, R., Schutzer, S. E., Ecker, D. J., Yang, T., Ramadoss, N. S., Robinson, W. H., Soloski, M. J., Eshoo, M. W., Aucott, J. N. 2020

  Abstract

  Lyme disease is a tick-borne infection caused by the bacteria Borrelia burgdorferi Current diagnosis of early Lyme disease relies heavily on clinical criteria, including the presence of an erythema migrans rash. The sensitivity of current gold-standard diagnostic tests relies upon antibody formation, which is typically delayed and thus of limited utility in early infection. We conducted a study of blood and skin biopsy specimens from 57 patients with a clinical diagnosis of erythema migrans. Samples collected at the time of diagnosis were analyzed using an ultra-sensitive, PCR-based assay employing an isothermal amplification step and multiple primers. In 75.4% of patients, we directly detected one or more B. burgdorferi genotypes in the skin. Two-tier testing showed that 20 (46.5%) of those found to be PCR positive remained serologically negative at both acute and convalescent time points. Multiple genotypes were found in 3 (8%) of those where a specific genotype could be identified. The 13 participants who lacked PCR and serologic evidence for exposure to B. burgdorferi could be differentiated as a group from PCR positive participants by their levels of several immune markers as well as by clinical descriptors such as their number of acute symptoms and the pattern of their erythema migrans rash. These results suggest that within a Mid-Atlantic cohort, patient subgroups can be identified using PCR-based direct detection approaches. This may be particularly useful in future research such as vaccine trials and public health surveillance of tick-borne disease patterns.

  View details for DOI 10.1128/JCM.00615-20

  View details for PubMedID 33087434

• Vitamin D Polygenetic Risk Score and the Association with RA Autoantibodies Among First-Degree Relatives of RA Subjects Bemis, E., Young, K., Seifert, J., Feser, M., Deane, K. D., Demoruelle, M., O'Dell, J., Weisman, M., Gregersen, P., Keating, R., Robinson, W., Buckner, J., Langefeld, C., Guthridge, J., James, J., Holers, V., Norris, J. WILEY. 2020

  View details for Web of Science ID 000587568502028

• Bacterial Families Lachnospiraceae/Ruminococcaceae Are Immunologically Targeted in Individuals At-risk for RA and a Specific Strain Is Arthritogenic in Monocultured Gnotobiotic Mice Chriswell, M., Seifert, J., Blum, L., Bloom, M., Feser, M., Demoruelle, M., Norris, J., Deane, K. D., James, E., Buckner, J., Robinson, W., Holers, V., Kuhn, K. WILEY. 2020

  View details for Web of Science ID 000587568502016

• Factors Associated with an Increased Risk of Imminent Rheumatoid Arthritis in ACPA plus Individuals Kim, H., Peterson, R., Firestein, G., Boyle, D., Buckner, J., Posso, S., James, E., Robinson, W., Arbet, J., Moss, L., Gilmore, R., Hartje, L., Barzideh, S., Seifert, J., Rao, N., Baribaud, F., Nagpal, S., Johnson, A., Holers, V., Deane, K. D. WILEY. 2020

  View details for Web of Science ID 000587568505224

• Higher Baseline Fine-Specificity ACPAs Predict Greater Treatment Response with Abatacept plus MTX versus MTX Monotherapy in Seropositive RA: A Post Hoc Analysis Robinson, W., Wu, C., Hu, S., Connolly, S., Mukherjee, S. WILEY. 2020

  View details for Web of Science ID 000587568501254

• Fine Specificity Anti-Citrullinated Protein Antibodies as Biomarkers for Prediction of Incident Rheumatoid Arthritis-Associated Interstitial Lung Disease Kronzer, V., Huang, W., Dellaripa, P., Huang, S., Feathers, V., Lu, B., Iannaccone, C., Gill, R., Hatabu, H., Nishino, M., Crowson, C., Davis, J., Robinson, W., Rughwani, T., Sokolove, J., Weinblatt, M., Shadick, N., Doyle, T. J., Sparks, J. WILEY. 2020

  View details for Web of Science ID 000587568501002

• Ultrasound in Knee Osteoarthritis: Reader Performance, Sonographic Features, and Correlation with Radiographic Findings Nwawka, O., Lee, S., Lin, B., Ibrahim, S., Brantner, C., Sculco, P., Parks, M., Figgie, M., Donlin, L., Orange, D., Mirza, S., Cross, M., Rodriguez, J., Sculco, T., Otero, M., Robinson, W., Goodman, S., Mehta, B. WILEY. 2020

  View details for Web of Science ID 000587568505045

• First-in-human phase Ib study of ATRC-101, an engineered version of a patient-derived antibody targeting a tumor-restricted ribonucleoprotein complex. Benjamin, J., Higgins, N., Millward, C., Defalco, J., Manning-Bog, A., Aydin, I. T., Zhang, D., Lippow, S., Scholz, A., Leung, Y., Zhu, Y., Sedello, A., Rogers, Z., Dhawan, I., Emerling, D., Robinson, W. H., Serafini, T., Greenberg, N. LIPPINCOTT WILLIAMS & WILKINS. 2020

  View details for Web of Science ID 000560368309083

• Erratum for Research Article "Synovial fibroblast-neutrophil interactions promote pathogenic adaptive immunity in rheumatoid arthritis" by C. Carmona-Rivera, P. M. Carlucci, E. Moore, N. Lingampalli, H. Uchtenhagen, E. James, Y. Liu, K. L. Bicker, H. Wahamma, V. Hoffman, A. I. Catrina, P. Thompson, J. H. Buckner, W. H. Robinson, D. A. Fox, M. J. Kaplan. Science immunology Carmona-Rivera, C., Carlucci, P. M., Moore, E., Lingampalli, N., Uchtenhagen, H., James, E., Liu, Y., Bicker, K. L., Wahamaa, H., Hoffmann, V., Catrina, A. I., Thompson, P. R., Buckner, J. H., Robinson, W. H., Fox, D. A., Kaplan, M. J. 2020; 5 (43)

  View details for DOI 10.1126/sciimmunol.aaz9319

  View details for PubMedID 32005681

  View details for PubMedCentralID PMC7228189

• Elevated anti-citrullinated protein antibodies prior to rheumatoid arthritis diagnosis and risks for chronic obstructive pulmonary disease or asthma. Arthritis care & research Zaccardelli, A., Liu, X., Ford, J. A., Cui, J., Lu, B., Chu, S. H., Schur, P. H., Speyer, C. B., Costenbader, K. H., Robinson, W. H., Sokolove, J., Karlson, E. W., Camargo, C. A., Sparks, J. A. 2020

  Abstract

  OBJECTIVE: To investigate elevation of anti-citrullinated protein antibodies (ACPA) before RA diagnosis and risks for chronic obstructive pulmonary disease (COPD) or asthma.METHODS: We performed a matched cohort study nested within the Nurses' Health Studies among women who donated blood. Women with incident RA after blood draw (self-reported then confirmed by medical records) were each matched to three controls by age, cohort, year, and menopausal factors. Pre-RA ACPA+ was defined as >99th percentile of control distribution by a research assay or by CCP2 in a subset. Incident COPD and asthma after index date (date of blood draw) were identified by questionnaires. Cox regression estimated HRs for incident COPD or asthma (in separate analyses) associated with pre-RA, pre-RA ACPA+, or pre-RA ACPA- phenotypes each compared to their matched non-RA controls.RESULTS: We analyzed 283 pre-RA women and 842 controls; blood was donated mean of 9.7 years (SD 5.8) before RA diagnosis. Fifty-nine women (20.8%) were pre-RA ACPA+. There were 107 cases of incident COPD and 105 incident asthma cases during 21,489 person-years of follow-up. Pre-RA ACPA+ was associated with increased COPD risk (HR 3.04, 95%CI 1.33,7.00) after adjusting for covariates including smoking pack-years. Pre-RA ACPA+ had a HR for asthma of 1.74 (multivariable 95%CI 0.72,4.24), similar to the risk of asthma for pre-RA ACPA- (HR 1.65, 95%CI 1.11,2.46).CONCLUSION: Women with elevated ACPA before RA diagnosis had increased risk for developing COPD compared to controls. Women who later developed RA were more likely to develop asthma, regardless of pre-RA ACPA status.

  View details for DOI 10.1002/acr.24140

  View details for PubMedID 31961487

• B cells in rheumatoid arthritis synovial tissues encode focused antibody repertoires that include antibodies that stimulate macrophage TNF-α production. Clinical immunology (Orlando, Fla.) Elliott, S. E., Kongpachith, S. n., Lingampalli, N. n., Adamska, J. Z., Cannon, B. J., Blum, L. K., Bloom, M. S., Henkel, M. n., McGeachy, M. J., Moreland, L. W., Robinson, W. H. 2020: 108360

  Abstract

  Rheumatoid arthritis (RA) is characterized by the production of anti-citrullinated protein antibodies (ACPAs). To gain insights into the relationship between ACPA-expressing B cells in peripheral blood (PB) and synovial tissue (ST), we sequenced the B cell repertoire in paired PB and ST samples from five individuals with established, ACPA+ RA. Bioinformatics analysis of paired heavy and light chain sequences revealed clonally-related family members shared between PB and ST. ST-derived antibody repertoires exhibited reduced diversity and increased normalized clonal family size compared to PB-derived repertoires. Functional characterization showed that seven recombinant antibodies (rAbs) expressed from subject-derived sequences from both compartments bound citrullinated antigens and immune complexes (ICs) formed using one ST-derived rAb stimulated macrophage TNF-α production. Our findings demonstrate B cell trafficking between PB and ST in subjects with RA and ST repertoires include B cells that encode ACPA capable of forming ICs that stimulate cellular responses implicated in RA pathogenesis.

  View details for DOI 10.1016/j.clim.2020.108360

  View details for PubMedID 32035179

• Soaking of Autologous Tendon Grafts in Vancomycin Before Implantation Does Not Lead to Tenocyte Cytotoxicity. The American journal of sports medicine Xiao, M. n., Leonardi, E. A., Sharpe, O. n., Sherman, S. L., Safran, M. R., Robinson, W. H., Abrams, G. D. 2020: 363546520951815

  Abstract

  Surgical site infections (SSIs) after anterior cruciate ligament (ACL) reconstruction procedures are an unfortunate complication. Soaking grafts in vancomycin before implantation has been reported to reduce the incidence of postoperative SSI after ACL reconstruction. There is potential for vancomycin to compromise graft integrity because of tenocyte toxicity.To examine the in vitro toxicity of varying doses of vancomycin on human tenocytes.Controlled laboratory study.Human patellar tenocytes were isolated and expanded in vitro. Tenocytes in culture were exposed to vancomycin at 5 different concentrations (400, 1600, 3200, 6400, and 12,800 μg/mL) and 3 time intervals (2, 6, and 24 hours). The control for all series was tenocyte exposure to only culture medium for each time interval. After treatment, a 10% Cell Counting Kit-8 solution in cellular growth medium was applied to the cells to examine cytotoxicity. A live/dead assay was used to assess tenocyte viability through fluorescence microscopy and flow cytometry. Results were analyzed statistically using multivariable logistic regression models with Tukey honest significant difference post hoc tests.Vancomycin did not cause significant changes in tenocyte viability after 2 and 6 hours of incubation at any concentration between 0 and 12,800 µg/mL. Incubation with vancomycin for 24 hours led to a significant decrease in cell viability at higher concentrations.Tenocytes derived from human patellar tendons exposed to relatively high concentrations of vancomycin for short periods of time do not demonstrate significant cell death and toxicity.Exposing tendons to vancomycin for a short period of time, such as before ACL reconstruction, is not likely to cause tenocyte toxicity because of vancomycin administration.

  View details for DOI 10.1177/0363546520951815

  View details for PubMedID 32898431

• CD52 Is Elevated on B cells of SLE Patients and Regulates B Cell Function. Frontiers in immunology Bhamidipati, K. n., Silberstein, J. L., Chaichian, Y. n., Baker, M. C., Lanz, T. V., Zia, A. n., Rasheed, Y. S., Cochran, J. R., Robinson, W. H. 2020; 11: 626820

  Abstract

  Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by B cell dysregulation and breaks in tolerance that lead to the production of pathogenic autoantibodies. We performed single-cell RNA sequencing of B cells from healthy donors and individuals with SLE which revealed upregulated CD52 expression in SLE patients. We further demonstrate that SLE patients exhibit significantly increased levels of B cell surface CD52 expression and plasma soluble CD52, and levels of soluble CD52 positively correlate with measures of lupus disease activity. Using CD52-deficient JeKo-1 cells, we show that cells lacking surface CD52 expression are hyperresponsive to B cell receptor (BCR) signaling, suggesting an inhibitory role for the surface-bound protein. In healthy donor B cells, antigen-specific BCR-activation initiated CD52 cleavage in a phospholipase C dependent manner, significantly reducing cell surface levels. Experiments with recombinant CD52-Fc showed that soluble CD52 inhibits BCR signaling in a manner partially-dependent on Siglec-10. Moreover, incubation of unstimulated B cells with CD52-Fc resulted in the reduction of surface immunoglobulin and CXCR5. Prolonged incubation of B cells with CD52 resulted in the expansion of IgD+IgMlo anergic B cells. In summary, our findings suggest that CD52 functions as a homeostatic protein on B cells, by inhibiting responses to BCR signaling. Further, our data demonstrate that CD52 is cleaved from the B cell surface upon antigen engagement, and can suppress B cell function in an autocrine and paracrine manner. We propose that increased expression of CD52 by B cells in SLE represents a homeostatic mechanism to suppress B cell hyperactivity.

  View details for DOI 10.3389/fimmu.2020.626820

  View details for PubMedID 33658999

  View details for PubMedCentralID PMC7917337

• PRIME Time in Rheumatoid Arthritis. The New England journal of medicine Gravallese, E. M., Robinson, W. H. 2020; 383 (3): 278–79

  View details for DOI 10.1056/NEJMe2018218

  View details for PubMedID 32668119

• Osteoarthritis risk is reduced after treatment with ticagrelor compared to clopidogrel: a propensity score matching analysis. Arthritis & rheumatology (Hoboken, N.J.) Baker, M. C., Weng, Y. n., William, R. H., Ahuja, N. n., Rohatgi, N. n. 2020

  Abstract

  Osteoarthritis (OA) is a common cause of joint pain and disability, and effective treatments are lacking. Extracellular adenosine has anti-inflammatory effects and can prevent and treat OA in animal models. Ticagrelor and clopidogrel are both used in patients with coronary artery disease, but only ticagrelor increases extracellular adenosine. The aim of this study was to determine whether treatment with ticagrelor was associated with a lower risk of OA.We conducted a 1:2 propensity score matching analysis using the Optum Clinformatics™ Data Mart from 2011 to 2017. We included patients who received either ticagrelor or clopidogrel for at least 90 days and excluded those with a prior diagnosis of OA or inflammatory arthritis. OA was identified using International Classification of Diseases codes. The primary outcome was the time to diagnosis of OA after treatment with ticagrelor versus clopidogrel.Our propensity score matched cohort consisted of 7,007 ticagrelor-treated patients and 14,014 clopidogrel-treated patients, with a median number of days on treatment of 287 and 284 respectively. For both groups, the mean age was 64 years, and 73% of the patients were male. Multivariate Cox-regression analysis estimated a hazard ratio of 0.71 (95% CI 0.64-0.79, p<0.001) for developing OA after treatment with ticagrelor compared to clopidogrel.Treatment with ticagrelor was associated with a 29% lower risk of developing OA compared to clopidogrel over five years of follow-up. We hypothesize that the reduction in OA seen in patients who received ticagrelor may in part be due to increased extracellular adenosine.

  View details for DOI 10.1002/art.41412

  View details for PubMedID 32564514

• Autoantibodies against central nervous system antigens in a subset of B cell-dominant multiple sclerosis patients. Proceedings of the National Academy of Sciences of the United States of America Kuerten, S. n., Lanz, T. V., Lingampalli, N. n., Lahey, L. J., Kleinschnitz, C. n., Mäurer, M. n., Schroeter, M. n., Braune, S. n., Ziemssen, T. n., Ho, P. P., Robinson, W. H., Steinman, L. n. 2020

  Abstract

  Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS), with characteristic inflammatory lesions and demyelination. The clinical benefit of cell-depleting therapies targeting CD20 has emphasized the role of B cells and autoantibodies in MS pathogenesis. We previously introduced an enzyme-linked immunospot spot (ELISpot)-based assay to measure CNS antigen-specific B cells in the blood of MS patients and demonstrated its usefulness as a predictive biomarker for disease activity in measuring the successful outcome of disease-modifying therapies (DMTs). Here we used a planar protein array to investigate CNS-reactive antibodies in the serum of MS patients as well as in B cell culture supernatants after polyclonal stimulation. Anti-CNS antibody reactivity was evident in the sera of the MS cohort, and the antibodies bound a heterogeneous set of molecules, including myelin, axonal cytoskeleton, and ion channel antigens, in individual patients. Immunoglobulin reactivity in supernatants of stimulated B cells was directed against a broad range of CNS antigens. A group of MS patients with a highly active B cell component was identified by the ELISpot assay. Those antibody reactivities remained stable over time. These assays with protein arrays identify MS patients with a highly active B cell population with antibodies directed against a swathe of CNS proteins.

  View details for DOI 10.1073/pnas.2011249117

  View details for PubMedID 32817492

• Protein phosphatase magnesium-dependent 1A induces inflammation in rheumatoid arthritis. Biochemical and biophysical research communications Lee, B., Song, Y. S., Rhodes, C., Goh, T. S., Roh, J. S., Jeong, H., Park, J., Lee, H., Lee, S., Kim, S., Kim, M., Lee, S., Sohn, D. H., Robinson, W. H. 2019

  Abstract

  Rheumatoid arthritis (RA) is a highly inflammatory autoimmune disease. Although proinflammatory cytokines, including tumor necrosis factor (TNF) and interleukin (IL)-6, play a key role in the pathogenesis of RA, the causes of chronic inflammation are not fully understood. Here, we report that protein phosphatase magnesium-dependent 1A (PPM1A) levels were increased in RA synovial fluid compared with osteoarthritis (OA) synovial fluid and positively correlated with TNF levels. In addition, PPM1A expression was increased in synovial tissue from RA patients and joint tissue from a mouse model of arthritis. Finally, extracellular PPM1A induced inflammation by stimulating macrophages to produce TNF through toll-like receptor 4 (TLR4) and myeloid differentiation primary response protein 88 (MyD88) signaling pathway. Our findings suggest that extracellular PPM1A may contribute to the pathogenesis of RA by functioning as a damage-associated molecular pattern (DAMP) to induce inflammation.

  View details for DOI 10.1016/j.bbrc.2019.11.112

  View details for PubMedID 31791585

• The intracellular protein phosphatase magnesium-dependent 1A negatively regulates osteoclast commitment. Arthritis & rheumatology (Hoboken, N.J.) Kwon, O. C., Choi, B., Lee, E., Park, J., Lee, E., Kim, E., Kim, S., Shin, M., Kim, T., Hong, S., Lee, C., Yoo, B., William, R. H., Kim, Y., Chang, E. 2019

  Abstract

  OBJECTIVE: Increased protein phosphatase magnesium-dependent 1A (PPM1A) levels in patients with ankylosing spondylitis regulate osteoblast differentiation in bony ankylosis; however, the potential mechanisms that regulate osteoclast (OC) differentiation in relation to abnormal bone formation remain unclear. Therefore, we generated conditional gene knockout (PPM1Afl/fl ;LysM-Cre) mice and evaluated their bone phenotype.METHODS: The bone phenotypes of LysM-Cre (n=6) and PPM1Afl/fl ;LysM-Cre mice (n=6) were assessed via micro-computed tomography. OC differentiation was induced by culturing bone marrow-derived macrophages in the presence of the RANKL and M-CSF, and was evaluated by counting tartrate-resistant acid phosphatase-positive multinucleated cells. The mRNA expressions of PPM1A, RANK and OC-specific genes were examined by quantitative real-time PCR, and protein levels were determined using Western blotting. Surface RANK expression was analysed by fluorescent flow cytometry.RESULTS: The PPM1Afl/fl ;LysM-Cre mice displayed reduced bone mass (p<0.001) and increased OC differentiation (p<0.001) and OC-specific gene expression (p<0.05) compared with their LysM-Cre littermates. Mechanistically, reduced PPM1A function in OC precursors in PPM1Afl/fl ;LysM-Cre mice induced OC lineage commitment by up-regulating RANK expression (p<0.01) via p38 MAPK activation in response to M-CSF. PPM1A expression in macrophages was decreased by TLR4 activation (p<0.05). The ankylosing spondylitis disease activity score was negatively correlated with the expression of PPM1A in peripheral blood mononuclear cells from Ax SpA patients (gamma=-0.7072, p<0.0001).CONCLUSION: The loss of PPM1A function in OC precursors driven by inflammatory signals contributes to OC lineage commitment and differentiation by elevating RANK expression, reflecting a potential role of PPM1A in dynamic bone metabolism in Ax SpA.

  View details for DOI 10.1002/art.41180

  View details for PubMedID 31762216

• Asthma and elevation of anti-citrullinated protein antibodies prior to the onset of rheumatoid arthritis. Arthritis research & therapy Zaccardelli, A., Liu, X., Ford, J. A., Cui, J., Lu, B., Chu, S. H., Schur, P. H., Speyer, C. B., Costenbader, K. H., Robinson, W. H., Sokolove, J., Karlson, E. W., Camargo, C. A., Sparks, J. A. 2019; 21 (1): 246

  Abstract

  BACKGROUND: Anti-citrullinated protein antibodies (ACPA) are central to rheumatoid arthritis (RA) pathogenesis and may develop at inflamed mucosa. We investigated whether asthma, a disease of airway mucosal inflammation, was associated with elevated ACPA before RA diagnosis.METHODS: We performed a nested case-control study among women in two prospective cohorts, the Nurses' Health Study (NHS; 1976-2014) and NHSII (1989-2015). Blood was obtained on a subset (NHS: 1989-1990; NHSII: 1996-1999). Cases met 1987 ACR or 2010 ACR/EULAR RA criteria by medical record review and were classified as seropositive (ACPA+ or rheumatoid factor positivity) or seronegative by clinical laboratory testing at diagnosis. We identified RA cases with blood drawn before the date of RA diagnosis (index date), matching each to three controls by age, cohort, year, time from blood draw to index date, and menopause. Pre-RA ACPA elevation for cases was defined as >99th percentile of the control distribution on a research assay composed of autoantibodies targeting citrullinated protein epitopes or positivity on the second-generation commercial assay for cyclic citrullinated peptide. Asthma status and covariates were obtained through biennial questionnaires before blood draw. Conditional logistic regression estimated ORs and 95%CIs for RA by pre-RA ACPA and clinical serostatus, adjusted for matching factors, smoking pack-years, passive smoking, and body mass index (BMI).RESULTS: We identified 284 incident RA cases and 849 matched controls; mean age at the index date was 61.2years (SD 10.1). Blood was drawn 9.7years (mean; SD 5.8) before the index date. We identified 96 (33.8%) RA cases with elevated pre-RA ACPA. At blood draw, 17.7% of pre-RA ACPA+ cases and 6.3% of matched controls (p=0.0008) reported clinician-diagnosed asthma. After adjusting for matching factors, smoking pack-years, passive smoking, and BMI, asthma was significantly associated with pre-RA ACPA+ RA (OR 3.57, 95%CI 1.58,8.04). Asthma was not associated with overall RA (OR 1.45, 95%CI 0.91,2.31), but was significantly associated with seropositive RA (OR 1.79, 95%CI 1.01,3.18).CONCLUSIONS: Asthma was strongly associated with ACPA elevation in blood drawn prior to RA diagnosis, independent of smoking. Chronic mucosal airway inflammation may contribute to ACPA development and RA pathogenesis.

  View details for DOI 10.1186/s13075-019-2035-3

  View details for PubMedID 31753003

• ATRC-101: A first-in-class engineered fully human monoclonal antibody that targets a tumor-restricted ribonucleoprotein complex Del-alco, J., Emerling, D., Manning-Bog, A., Baia, G., Lippow, S., Scholz, A., Zhu, Y., Cavet, G., Volkmuth, W., Dhawan, I., Benjamin, J., Robinson, W., Serafini, T., Greenberg, N. BMC. 2019

  View details for Web of Science ID 000496473200274

• Antibody Repertoire Sequencing, Antigen Array Analysis, and Cytokine Profiling of Blood from Individuals at High-risk for RA Reveals Candidate Immunoglobulin V Genes, ACPA, and Cytokines That May Promote the Transition to Arthritis Iyer, R., Zia, A., Bloom, M., Nagpal, S., Rao, N., Baribaud, F., Vratsanos, G., Wang, W., Firestein, G., Boyle, D., Buckner, J., Holers, V., Deane, K., Robinson, W. WILEY. 2019

  View details for Web of Science ID 000507466900483

• Differences in the Phenotypic Landscape and Antigen Specificity of CD4+T Cells Are Present in CCP plus Subjects Before the Onset of Rheumatoid Arthritis Muir, V., Rims, C., Deane, K., Carlin, J., Posso, S., Nagpal, S., Rao, N., Baribaud, F., Vratsanos, G., Robinson, W., Firestein, G., Holers, V., Linsley, P., James, E., Buckner, J. WILEY. 2019

  View details for Web of Science ID 000507466905048

• Elevation of Anti-Citrullinated Protein Antibodies Prior to Rheumatoid Arthritis Onset and Risks for Developing Chronic Obstructive Pulmonary Disease or Asthma Zaccardelli, A., Liu, X., Ford, J., Cui, J., Lu, B., Chu, S. H., Schur, P., Speyer, C., Costenbader, K., Robinson, W., Sokolove, J., Karlson, E., Camargo, C., Sparks, J. WILEY. 2019

  View details for Web of Science ID 000507466901264

• Differential Methylation of Peripheral Blood Adaptive Immune Cells in Individuals at High Risk for RA and with Early RA Compared with Controls Identifies Pathways Important in Transition to Arthritis Ai, R., Boyle, D., Hammaker, D., Deane, K., Holers, V., Matti, A., Robinson, W., Buckner, J., Rao, N., Baribaud, F., Vratsanos, G., Nagpal, S., Wang, W., Firestein, G. WILEY. 2019

  View details for Web of Science ID 000507466904502

• Clinical and Biomarker Factor Associations with Symptoms and Future Development of RA: TIP-RA Collective Deane, K., Nagpal, S., Holers, V., Firestein, G., Boyle, D., Seifert, J., Buckner, J., James, E., Moss, L., Barzideh, S., Posso, S., Robinson, W., Polinski, K., Gilmore, R., Baribaud, F., Rao, N., Vratsanos, G. WILEY. 2019

  View details for Web of Science ID 000507466900495

• Associations Between Circulating Cytokines and Incident Inflammatory Arthritis in an Anti-citrullinated Protein Antibody Positive Population Polinski, K., Bemis, E., Demoruelle, M., Feser, M., Moss, L., Seifert, J., Clare-Salzler, M., Robinson, W., Holers, V., Deane, K., Norris, J. WILEY. 2019

  View details for Web of Science ID 000507466900176

• Neutrophil extracellular traps, B cells, and type I interferons contribute to immune dysregulation in hidradenitis suppurativa. Science translational medicine Byrd, A. S., Carmona-Rivera, C., O'Neil, L. J., Carlucci, P. M., Cisar, C., Rosenberg, A. Z., Kerns, M. L., Caffrey, J. A., Milner, S. M., Sacks, J. M., Aliu, O., Broderick, K. P., Reichner, J. S., Miller, L. S., Kang, S., Robinson, W. H., Okoye, G. A., Kaplan, M. J. 2019; 11 (508)

  Abstract

  Hidradenitis suppurativa (HS), also known as acne inversa, is an incapacitating skin disorder of unknown etiology manifested as abscess-like nodules and boils resulting in fistulas and tissue scarring as it progresses. Given that neutrophils are the predominant leukocyte infiltrate in HS lesions, the role of neutrophil extracellular traps (NETs) in the induction of local and systemic immune dysregulation in this disease was examined. Immunofluorescence microscopy was performed in HS lesions and detected the prominent presence of NETs. NET complexes correlated with disease severity, as measured by Hurley staging. Neutrophils from the peripheral blood of patients with HS peripheral also displayed enhanced spontaneous NET formation when compared to healthy control neutrophils. Sera from patients recognized antigens present in NETs and harbored increased antibodies reactive to citrullinated peptides. B cell dysregulation, as evidenced by elevated plasma cells and IgG, was observed in the circulation and skin from patients with HS. Peptidylarginine deiminases (PADs) 1 to 4, enzymes involved in citrullination, were differentially expressed in HS skin, when compared to controls, in association with enhanced tissue citrullination. NETs in HS skin coexisted with plasmacytoid dendritic cells, in association with a type I interferon (IFN) gene signature. Enhanced NET formation and immune responses to neutrophil and NET-related antigens may promote immune dysregulation and contribute to inflammation. This, along with evidence of up-regulation of the type I IFN pathway in HS skin, suggests that the innate immune system may play important pathogenic roles in this disease.

  View details for DOI 10.1126/scitranslmed.aav5908

  View details for PubMedID 31484788

• Antigen-specific tolerance to self-antigens in protein replacement therapy, gene therapy and autoimmunity. Current opinion in immunology Steinman, L., Ho, P. P., Robinson, W. H., Utz, P. J., Villoslada, P. 2019; 61: 46–53

  Abstract

  Trials of antigen-specific tolerance have been undertaken in the clinic for over fifty years and the results of these antigen-specific clinical trials are described in this review. Antigen-specific tolerization of the immune system in protein replacement therapy for hemophilia A is an accepted treatment. Clinical trials are ongoing for autoimmune conditions such as type 1 diabetes, multiple sclerosis, neuromyelitis optica, and rheumatoid arthritis with various antigen-specific strategies. Trials for tolerization in celiac disease aim for antigen specific tolerance to gluten, an environmental trigger, which may then halt the progression to autoimmunity targeting a self-antigen, tissue transglutaminase. Although many promising approaches have been demonstrated in pre-clinical models, this review will focus primarily on clinical trials of antigen-specific tolerance that have been taken to the clinic and with initial results reported in the peer reviewed literature. A separate article on approaches with CAR-T cells appears in this volume.

  View details for DOI 10.1016/j.coi.2019.07.011

  View details for PubMedID 31476445

• Platelet-Rich Plasma (PRP) From Older Males With Knee Osteoarthritis Depresses Chondrocyte Metabolism and Upregulates Inflammation JOURNAL OF ORTHOPAEDIC RESEARCH O'Donnell, C., Migliore, E., Grandi, F., Koltsov, J., Lingampalli, N., Cisar, C., Indelli, P. F., Sebastiano, V., Robinson, W. H., Bhutani, N., Chu, C. R. 2019; 37 (8): 1760–70

  View details for DOI 10.1002/jor.24322

  View details for Web of Science ID 000501249400010

• A Prospective Study of the Development of Inflammatory Arthritis in the Family Members of Indigenous North American People With Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Tanner, S., Dufault, B., Smolik, I., Meng, X., Anaparti, V., Hitchon, C., Robinson, D. B., Robinson, W., Sokolove, J., Lahey, L., Ferucci, E. D., El-Gabalawy, H. 2019

  View details for DOI 10.1002/art.40880

  View details for Web of Science ID 000479771200001

• Defining inflammatory cell states in rheumatoid arthritis joint synovial tissues by integrating single-cell transcriptomics and mass cytometry NATURE IMMUNOLOGY Zhang, F., Wei, K., Slowikowski, K., Fonseka, C. Y., Rao, D. A., Kelly, S., Goodman, S. M., Tabechian, D., Hughes, L. B., Salomon-Escoto, K., Watts, G. M., Jonsson, A., Rangel-Moreno, J., Meednu, N., Rozo, C., Apruzzese, W., Eisenhaure, T. M., Lieb, D. J., Boyle, D. L., Mandelin, A. M., Boyce, B. F., DiCarlo, E., Gravallese, E. M., Gregersen, P. K., Moreland, L., Firestein, G. S., Hacohen, N., Nusbaum, C., Lederer, J. A., Perlman, H., Pitzalis, C., Filer, A., Holers, V., Bykerk, V. P., Donlin, L. T., Anolik, J. H., Brenner, M. B., Raychaudhuri, S., Albrecht, J., Bridges, S., Buckley, C. D., Buckner, J. H., Dolan, J., Guthridge, J. M., Gutierrez-Arcelus, M., Ivashkiv, L. B., James, E. A., James, J. A., Keegan, J., Lee, Y. C., McGeachy, M. J., McNamara, M. A., Mears, J. R., Mizoguchi, F., Nguyen, J. P., Noma, A., Orange, D. E., Rohani-Pichavant, M., Ritchlin, C., Robinson, W. H., Seshadri, A., Sutherby, D., Seifert, J., Turner, J. D., Utz, P. J., Accelerating Medicines Partnersh 2019; 20 (7): 928-+

  Abstract

  To define the cell populations that drive joint inflammation in rheumatoid arthritis (RA), we applied single-cell RNA sequencing (scRNA-seq), mass cytometry, bulk RNA sequencing (RNA-seq) and flow cytometry to T cells, B cells, monocytes, and fibroblasts from 51 samples of synovial tissue from patients with RA or osteoarthritis (OA). Utilizing an integrated strategy based on canonical correlation analysis of 5,265 scRNA-seq profiles, we identified 18 unique cell populations. Combining mass cytometry and transcriptomics revealed cell states expanded in RA synovia: THY1(CD90)+HLA-DRAhi sublining fibroblasts, IL1B+ pro-inflammatory monocytes, ITGAX+TBX21+ autoimmune-associated B cells and PDCD1+ peripheral helper T (TPH) cells and follicular helper T (TFH) cells. We defined distinct subsets of CD8+ T cells characterized by GZMK+, GZMB+, and GNLY+ phenotypes. We mapped inflammatory mediators to their source cell populations; for example, we attributed IL6 expression to THY1+HLA-DRAhi fibroblasts and IL1B production to pro-inflammatory monocytes. These populations are potentially key mediators of RA pathogenesis.

  View details for DOI 10.1038/s41590-019-0378-1

  View details for Web of Science ID 000471891900022

  View details for PubMedID 31061532

• Histologic and Transcriptional Evidence of Subclinical Synovial Inflammation in Patients With Rheumatoid Arthritis in Clinical Remission ARTHRITIS & RHEUMATOLOGY Orange, D. E., Agius, P., DiCarlo, E. F., Mirza, S. Z., Pannellini, T., Szymonifka, J., Jiang, C. S., Figgie, M. P., Frank, M. O., Robinson, W. H., Donlin, L. T., Rozo, C., Gravallese, E. M., Bykerk, V. P., Goodman, S. M. 2019; 71 (7): 1034–41

  View details for DOI 10.1002/art.40878

  View details for Web of Science ID 000472975100004

• ASSOCIATIONS OF BASELINE CLINICAL AND BIOMARKER FACTORS WITH SYMPTOMS AND FUTURE DEVELOPMENT OF CLINICALLY-APPARENT RHEUMATOID ARTHRITIS IN AN ACPA POSITIVE COHORT Deane, K. D., Nagpal, S., Rao, N., Baribaud, F., Vratsanos, G., James, E. A., Buckner, J. H., Firestein, G. S., Boyle, D. L., Posso, S., Robinson, W. H., Moss, L. K., Barzideh, S., Polinski, K., Ivanov, S., Seifert, J., Holers, V. BMJ PUBLISHING GROUP. 2019: 1105

  View details for DOI 10.1136/annrheumdis-2019-eular.1297

  View details for Web of Science ID 000472207103268

• ASTHMA AND ELEVATION OF ANTI-CITRULLINATED PROTEIN ANTIBODIES PRIOR TO THE ONSET OF RHEUMATOID ARTHRITIS Zaccardelli, A., Liu, X., Ford, J., Tedeschi, S., Cui, J., Lu, B., Chu, S., Schur, P., Speyer, C., Costenbader, K., Robinson, W., Sokolove, J., Camargo, C., Sparks, J. BMJ PUBLISHING GROUP. 2019: 642

  View details for DOI 10.1136/annrheumdis-2019-eular.3107

  View details for Web of Science ID 000472207102001

• Exogenous micro-RNA and antagomir modulate osteogenic gene expression in tenocytes EXPERIMENTAL CELL RESEARCH Xiao, M., Iglinski-Benjamin, K. C., Sharpe, O., Robinson, W. H., Abrams, G. D. 2019; 378 (2): 119–23

  View details for DOI 10.1016/j.yexcr.2019.03.008

  View details for Web of Science ID 000464772300001

• B cell checkpoints in autoimmune rheumatic diseases NATURE REVIEWS RHEUMATOLOGY Rubin, S. S., Bloom, M. S., Robinson, W. H. 2019; 15 (5): 303–15

  View details for DOI 10.1038/s41584-019-0211-0

  View details for Web of Science ID 000465571600012

• Platelet-Rich Plasma (PRP) from Older Males with Knee Osteoarthritis Depresses Chondrocyte Metabolism and Upregulates Inflammation. Journal of orthopaedic research : official publication of the Orthopaedic Research Society O'Donnell, C., Migliore, E., Grandi, F. C., Koltsov, J., Lingampalli, N., Cisar, C., Indelli, P. F., Sebastiano, V., Robinson, W. H., Bhutani, N., Chu, C. R. 2019

  Abstract

  There is intense clinical interest in the potential effects of platelet-rich plasma (PRP) for the treatment of osteoarthritis (OA). This study tested the hypotheses that (1) 'lower' levels of the inflammatory mediators (IM) interleukin-1-beta (IL-1beta) and tumor-necrosis-factor-alpha (TNF-alpha), and (2) 'higher' levels of the growth factors (GF) insulin-like-growth-factor-1 and transforming-growth-factor-beta-1 within leukocyte-poor PRP correlate with more favorable chondrocyte and macrophage responses in vitro. Samples were collected from ten 'healthy' young male (23-33 years old) human subjects (H-PRP) and nine older (62-85 years old) male patients with severe knee OA (OA-PRP). The samples were separated into groups of 'high' or 'low' levels of IM and GF based on multiplex cytokine and ELISA data. Three-dimensional (3D) alginate bead chondrocyte cultures and monocyte-derived macrophage cultures were treated with 10% PRP from donors in different groups. Gene expression was analyzed by qPCR. Contrary to our hypotheses, the effect of PRP on chondrocytes and macrophages was mainly influenced by the age and disease status of the PRP donor as opposed to the IM or GF groupings. While H-PRP showed similar effects on expression of chondrogenic markers (Col2a1 and Sox9) as the negative control group (p>0.05), OA-PRP decreased chondrocyte expression of Col2a1 and Sox-9 mRNA by 40% and 30%, respectively (Col2a1, p=0.015; Sox9, p=0.037). OA-PRP also upregulated TNF-alpha and MMP-9 (p<0.001) gene expression in macrophages while H-PRP did not. This data suggests that PRP from older individuals with OA contain factors that may suppress chondrocyte matrix synthesis and promote macrophage inflammation in vitro. This article is protected by copyright. All rights reserved.

  View details for PubMedID 31042308

• B cell checkpoints in autoimmune rheumatic diseases. Nature reviews. Rheumatology Rubin, S. J., Bloom, M. S., Robinson, W. H. 2019

  Abstract

  B cells have important functions in the pathogenesis of autoimmune diseases, including autoimmune rheumatic diseases. In addition to producing autoantibodies, B cells contribute to autoimmunity by serving as professional antigen-presenting cells (APCs), producing cytokines, and through additional mechanisms. B cell activation and effector functions are regulated byimmune checkpoints, including both activating and inhibitory checkpoint receptors that contribute to the regulation of B cell tolerance, activation, antigen presentation, T cell help, classswitching, antibody production and cytokine production. The various activating checkpoint receptors include B cell activating receptors that engage with cognate receptors on T cells or other cells, as well as Toll-like receptors that can provide dual stimulation to B cells via co-engagement with the B cell receptor. Furthermore, various inhibitory checkpoint receptors, including B cell inhibitory receptors, have important functions in regulating B cell development, activation and effector functions. Therapeutically targeting B cell checkpoints represents a promising strategy for the treatment of a variety of autoimmune rheumatic diseases.

  View details for PubMedID 30967621

• Latent autoimmunity across disease-specific boundaries in at-risk first-degree relatives of SLE and RA patients. EBioMedicine James, J. A., Chen, H., Young, K. A., Bemis, E. A., Seifert, J., Bourn, R. L., Deane, K. D., Demoruelle, M. K., Feser, M., O'Dell, J. R., Weisman, M. H., Keating, R. M., Gaffney, P. M., Kelly, J. A., Langefeld, C. D., Harley, J. B., Robinson, W., Hafler, D. A., O'Connor, K. C., Buckner, J., Guthridge, J. M., Norris, J. M., Holers, V. M. 2019

  Abstract

  BACKGROUND: Autoimmune disease prevention requires tools to assess an individual's risk of developing a specific disease. One tool is disease-associated autoantibodies, which accumulate in an asymptomatic preclinical period. However, patients sometimes exhibit autoantibodies associated with a different disease classification. When and how these alternative autoantibodies first appear remain unknown. This cross-sectional study characterizes alternative autoimmunity, and associated genetic and environmental factors, in unaffected first-degree relatives (FDRs) of patients, who exhibit increased future risk for the same disease.METHODS: Samples (n = 1321) from disease-specific autoantibody-positive (aAb+) systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and type 1 diabetes (T1D) patients; and unaffected aAb+ and autoantibody-negative (aAb-) SLE and RA FDRs were tested for SLE, RA, and T1D aAbs, as well as anti-tissue transglutaminase, anti-cardiolipin and anti-thyroperoxidase. FDR SLE and RA genetic risk scores (GRS) were calculated.FINDINGS: Alternative autoimmunity occurred in SLE patients (56%) and FDRs (57·4%), RA patients (32·6%) and FDRs (34·8%), and T1D patients (43%). Expanded autoimmunity, defined as autoantibodies spanning at least two other diseases, occurred in 18·5% of SLE patients, 16·4% of SLE FDRs, 7·8% of RA patients, 5·3% of RA FDRs, and 10·8% of T1D patients. SLE FDRs were more likely to have alternative (odds ratio [OR] 2·44) and expanded (OR 3·27) autoimmunity than RA FDRs. Alternative and expanded autoimmunity were associated with several environmental exposures. Alternative autoimmunity was associated with a higher RA GRS in RA FDRs (OR 1·41), and a higher SLE GRS in aAb+ RA FDRs (OR 1·87), but not in SLE FDRs.INTERPRETATION: Autoimmunity commonly crosses disease-specific boundaries in systemic (RA, SLE) and organ-specific (T1D) autoimmune diseases. Alternative autoimmunity is more common in SLE FDRs than RA FDRs, and is influenced by genetic and environmental factors. These findings have substantial implications for preclinical disease pathogenesis and autoimmune disease prevention studies. FUND: NIH U01AI101981, R01AR051394, U19AI082714, P30AR053483, P30GM103510, U54GM104938, U01AI101934, R01AI024717, U01AI130830, I01BX001834, & U01HG008666.

  View details for PubMedID 30952617

• Latent autoimmunity across disease-specific boundaries in at-risk first-degree relatives of SLE and RA patients EBIOMEDICINE James, J. A., Chen, H., Young, K. A., Bemis, E. A., Seifert, J., Bourn, R. L., Deane, K. D., Demoruelle, M., Feser, M., O'Dell, J. R., Weisman, M. H., Keating, R. M., Gaffney, P. M., Kelly, J. A., Langefeld, C. D., Harley, J. B., Robinson, W., Hafler, D. A., O'Connor, K. C., Buckner, J., Guthridge, J. M., Norris, J. M., Holers, V. 2019; 42: 76–85

  View details for DOI 10.1016/j.ebiom.2019.03.063

  View details for Web of Science ID 000466175100027

• Discovery of Distinct Immune Phenotypes Using Machine Learning in Pulmonary Arterial Hypertension CIRCULATION RESEARCH Sweatt, A. J., Hedlin, H. K., Balasubramanian, V., Hsi, A., Blum, L. K., Robinson, W. H., Haddad, F., Hickey, P. M., Condliffe, R., Lawrie, A., Nicolls, M. R., Rabinovitch, M., Khatri, P., Zamanian, R. T. 2019; 124 (6): 904–19

  View details for DOI 10.1161/CIRCRESAHA.118.313911

  View details for Web of Science ID 000469341600015

• Direct Diagnostic Tests for Lyme Disease CLINICAL INFECTIOUS DISEASES Schutzer, S. E., Body, B. A., Boyle, J., Branson, B. M., Dattwyler, R. J., Fikrig, E., Gerald, N. J., Gomes-Solecki, M., Kintrup, M., Ledizet, M., Levin, A. E., Lewinski, M., Liotta, L. A., Marques, A., Mead, P. S., Mongodin, E. F., Pillai, S., Rao, P., Robinson, W. H., Roth, K. M., Schriefer, M. E., Slezak, T., Snyder, J. L., Steere, A. C., Witkowski, J., Wong, S. J., Branda, J. A. 2019; 68 (6): 1052–57

  View details for DOI 10.1093/cid/ciy614

  View details for Web of Science ID 000461136900026

• Prefoldin 5 and Anti-prefoldin 5 Antibodies as Biomarkers for Uveitis in Ankylosing Spondylitis. Frontiers in immunology Kwon, O. C., Lee, E. J., Lee, J. Y., Youn, J., Kim, T. H., Hong, S., Lee, C. K., Yoo, B., Robinson, W. H., Kim, Y. G. 2019; 10: 384

  Abstract

  Objective: Uveitis is the most common extra-articular manifestation of ankylosing spondylitis (AS), for which no diagnostic biomarkers have been identified. This study was conducted to identify biomarker for uveitis in AS. Methods: To identify autoantibodies associated with uveitis in AS, we performed human protein microarray analysis using sera derived from various autoimmune diseases and ELISA analysis of sera derived from AS and rheumatoid arthritis patients. In the curdlan-induced SKG mice model, ophthalmic examination was performed at week 8 post-immunization and histologic examination of the ocular lesions performed at week 16 post-immunization. Serum levels of target antibodies were assessed at various time-points. To evaluate the functional role of specific autoantibodies, an in vitro apoptosis assay using ARPE-19 cells was performed. Results: Reactivity against prefoldin subunit 5 (PFDN5) was identified in AS with uveitis. Levels of anti-PFDN5 antibodies and PFDN5 in sera from AS with uveitis patients were significantly higher than those in AS without uveitis. At week 8, half of curdlan-treated SKG mice developed anterior uveitis, while all of them developed histologically confirmed uveitis at week 16. The levels of anti-PFDN5 antibodies increased over time in the sera of curdlan-treated SKG mice along with increased expression of PFDN5 and apoptosis in the ocular lesions. Knockdown of PFDN5 in ARPE19 cells resulted in increased apoptosis, suggesting a protective role of PFDN5 against cell death in uveitis. Conclusion: AS patients with uveitis have increased levels of anti-PFDN5 antibodies, and our findings suggest that anti-PFDN5 antibodies could provide a biomarker for uveitis in AS.

  View details for DOI 10.3389/fimmu.2019.00384

  View details for PubMedID 30891043

  View details for PubMedCentralID PMC6411661

• Prefoldin 5 and Anti-prefoldin 5 Antibodies as Biomarkers for Uveitis in Ankylosing Spondylitis FRONTIERS IN IMMUNOLOGY Kwon, O., Lee, E., Lee, J., Youn, J., Kim, T., Hong, S., Lee, C., Yoo, B., Robinson, W. H., Kim, Y. 2019; 10

  View details for DOI 10.3389/fimmu.2019.00384

  View details for Web of Science ID 000460293400002

• Histologic and transcriptional evidence of subclinical synovial inflammation in rheumatoid arthritis patients in clinical remission. Arthritis & rheumatology (Hoboken, N.J.) Orange, D. E., Agius, P., DiCarlo, E. F., Mirza, S., Pannellini, T., Szymonifka, J., Jiang, C. S., Figgie, M. P., Frank, M. O., Robinson, W. H., Donlin, L. T., Rozo, C., Gravallese, E. M., Bykerk, V. P., Goodman, S. M. 2019

  Abstract

  INTRODUCTION: Patients with rheumatoid arthritis (RA) in clinical remission may have subclinical synovial inflammation. Here, we sought to determine the proportion of patients in remission or low disease activity at the time of arthroplasty that have histologic or transcriptional evidence of synovitis, as well as clinical features that distinguish patients with subclinical synovitis.METHODS: We compared disease activity scores with 28 joint counts (DAS28) to synovial histology features in 135 patients with RA undergoing arthroplasty. We also compared DAS28 to RNA-Seq data on a subset of 35 patients.RESULTS: 14% of patients met DAS28 criteria for clinical remission (DAS28<2.6) and another 15% met criteria for low disease activity (DAS28<3.2). Histologic analysis of synovium revealed synovitis in 27% and 31% of samples from patients in remission and low disease activity respectively. Patients with low disease activity (DAS28<3.2) and synovitis also exhibited increased CRP and anti-citrullinated peptide antibody (CCP) levels compared to those without synovitis. 183 genes were differentially expressed in synovium of patients with subclinical synovitis compared to those with low inflammatory synovium. 86% of these genes were also differentially expressed in synovium of patients who were clinically active (DAS28≥3.2).CONCLUSION: 31% of patients with low clinical disease activity exhibit histologic evidence of subclinical synovitis, which was associated with increased CRP and CCP levels. Synovial gene expression signatures of clinical synovitis are present in patients with subclinical synovitis. This article is protected by copyright. All rights reserved.

  View details for PubMedID 30835943

• Exogenous Micro-RNA and Antagomir Modulate Osteogenic Gene Expression in Tenocytes. Experimental cell research Xiao, M., Iglinski-Benjamin, K. C., Sharpe, O., Robinson, W. H., Abrams, G. D. 2019

  Abstract

  Tendinopathy is a common and disabling condition that is difficult to treat. The pathomolecular events behind tendinopathy remain uncertain. Micro-RNAs (miRNAs, miRs) are short non-coding RNAs that regulate gene expression and may play a role in tendinopathy development. Tenocytes were obtained from human patellar tendons in patients undergoing anterior cruciate ligament (ACL) reconstruction. Micro-RNA mimics and antagomirs for miR-30d, 26a, and 29a were separately transfected into tenocyte culture. Gene expression for scleraxis, collagen 1 alpha 1 (COL1A1), collagen 3 alpha 1 (COL3A1), interleukin-1-beta (IL-1beta), interleukin-6 (IL-6), bone morphogenic protein 2 (BMP2), bone morphogenic protein 12 (BMP12), and osteocalcin was determined for each miRNA mimic and antagomir transfection using real-time quantitative PCR (qPCR). The results showed that exogenous miR-29a downregulated BMP2 and BMP12, while miR-26a and miR-30d did not have a significant effect on tenocyte gene expression. These findings suggest miR-29a contributes to tendon homeostasis and can serve as a potential therapeutic target in treating tendinopathy.

  View details for PubMedID 30849310

• PREFERENTIAL BINDING TO AN UNEXPECTED EPITOPE OF A CHIMERIC RECOMBINANT PROTEINASE 3 VARIANT BY ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODIES Thompson, G., Moura, M., Nelson, D., Fussner, L., Hummel, A., Jenne, D., Fervenza, F., Hoffman, G., Kallenberg, C., Langford, C., McCune, J., Merkel, P., Monach, P., Seo, P., Spiera, R., St Clair, E., Ytterberg, S., Stone, J., Robinson, W., Pang, Y., Specks, U. OXFORD UNIV PRESS. 2019

  View details for Web of Science ID 000478085100222

• Affinity Maturation of the Anti-Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis. Arthritis & rheumatology (Hoboken, N.J.) Kongpachith, S., Lingampalli, N., Ju, C., Blum, L. K., Lu, D. R., Elliott, S. E., Mao, R., Robinson, W. H. 2019

  Abstract

  OBJECTIVE: Anti-citrullinated protein antibodies (ACPAs) are a hallmark of rheumatoid arthritis (RA). While epitope spreading of the serum ACPA response is believed to contribute to RA pathogenesis, little is understood regarding how this phenomenon occurs. This study was undertaken to analyze the antibody repertoires of individuals with RA to gain insight into the mechanisms leading to epitope spreading of the serum ACPA response in RA.METHODS: Plasmablasts from the blood of 6 RA patients were stained with citrullinated peptide tetramers to identify ACPA-producing B cells by flow cytometry. Plasmablasts were single-cell sorted and sequenced to obtain antibody repertoires. Sixty-nine antibodies were recombinantly expressed, and their anticitrulline reactivities were characterized using a cyclic citrullinated peptide enzyme-linked immuosorbent assay and synovial antigen arrays. Thirty-six mutated antibodies designed either to represent ancestral antibodies or to test paratope residues critical for binding, as determined from molecular modeling studies, were also tested for anticitrulline reactivities.RESULTS: Clonally related monoclonal ACPAs and their shared ancestral antibodies each exhibited differential reactivity against citrullinated antigens. Molecular modeling identified residues within the complementarity-determining region loops and framework regions predicted to be important for citrullinated antigen binding. Affinity maturation resulted in mutations of these key residues, which conferred binding to different citrullinated epitopes and/or increased polyreactivity to citrullinated epitopes.CONCLUSION: These results demonstrate that the different somatic hypermutations accumulated by clonally related B cells during affinity maturation alter the antibody paratope to mediate epitope spreading and polyreactivity of the ACPA response in RA, suggesting that these may be key properties that likely contribute to the pathogenicity of ACPAs.

  View details for PubMedID 30811898

• A prospective study of the development of inflammatory arthritis in the family members of Indigenous North American people with rheumatoid arthritis. Arthritis & rheumatology (Hoboken, N.J.) Tanner, S. n., Dufault, B. n., Smolik, I. n., Meng, X. n., Anaparti, V. n., Hitchon, C. n., Robinson, D. B., William, R. n., Sokolove, J. n., Lahey, L. n., Ferucci, E. D., El-Gabalawy, H. n. 2019

  Abstract

  We prospectively followed a cohort of unaffected relatives of Indigenous North American (INA) people with RA to define the incidence of inflammatory arthritis (IA) and autoantibody prevalence in this population.Unaffected relatives of INA people with RA from central Canada and Alaska were followed systematically from 2005 until 2017. Rheumatoid factor (RF) and anti-citrullinated protein antibodies (ACPA) were tested at every visit and a subset was tested for ACPA fine specificity using a custom multiplex assay. Multi-state models based on all available study visits were developed to determine the likelihood of transitioning between autoantibody states, or to IA.18/374 (4.8%) relatives developed seropositive IA after 4.7 (±2.4) years of follow-up, giving a transition rate of 9.2 cases per 1000 person-years. Thirty percent of those who developed IA were seronegative at baseline but all were seropositive at IA onset. Although 30% of ACPA/RF double seropositive individuals developed IA after 3.9 (±2.6) years, the majority did not develop IA. Multi-state modeling indicated a 71% and 68% likelihood of ACPA and RF seropositive states, respectively, reverting to seronegativity after five years, and a 39% likelihood of ACPA/RF double seropositive state becoming seronegative. Fine specificity testing demonstrated expansion of the ACPA repertoire prior to development of IA.Despite a high incidence of IA in this cohort of at-risk relatives of INA people with RA, a large proportion of autoantibody positive individuals do not develop IA and revert back to an autoantibody negative state. This article is protected by copyright. All rights reserved.

  View details for PubMedID 30861615

• Fibroblast-like synovial cell production of extra domain A fibronectin associates with inflammation in osteoarthritis. BMC rheumatology Kragstrup, T. W., Sohn, D. H., Lepus, C. M., Onuma, K., Wang, Q., Robinson, W. H., Sokolove, J. 2019; 3: 46

  Abstract

  Background: The pathophysiology of osteoarthritis (OA) involves wear and tear, and a state of low-grade inflammation. Tissue repair responses include transforming growth factor beta (TGFbeta)-induced myofibroblast production of extracellular matrix. Fibronectins are an essential part of the extracellular matrix, and injection of fibronectin fragments into rabbit joints is a previously established animal model of OA. Fibronectin containing the ED-A domain is currently being used as drug delivery target in the development of anti-inflammatory drugs (e.g. Dekavil).Methods: In this study, samples of synovial membrane were obtained from patients with knee OA undergoing joint replacement surgery. Immunostaining for ED-A fibronectin and the myofibroblast marker alpha smooth muscle actin (alphaSMA) was performed on fibroblast-like synovial cells (FLS) and synovial membranes. RAW 264.7 macrophages were incubated with recombinant ED-A fibronectin.Results: The staining of ED-A fibronectin in OA FLS was increased by TGFbeta but not by TNFalpha, lipopolysaccharide, or IL-6 (n=3). ED-A fibronectin co-stained with the myofibroblast marker alphaSMA in both the OA FLS (n=3) and in the OA synovial membranes (n=8). ED-A fibronectin staining was associated with both number of lining layer cells (rho=0.85 and p=0.011) and sublining cells (rho=0.88 and p=0.007) in the OA synovium (n=8), and co-distributed with TNFalpha (n=5). Recombinant ED-A fibronectin increased the production of TNFalpha by RAW 264.7 macrophages (n=3).Conclusions: The disease process in OA shares features with the chronic wound healing response. Our findings support utilizing ED-A fibronectin for drug delivery or therapeutic targeting to reduce pro-inflammatory responses in OA.

  View details for DOI 10.1186/s41927-019-0093-4

  View details for PubMedID 31819923

• Remote Activation of a Latent Epitope in an Autoantigen Decoded With Simulated B-Factors. Frontiers in immunology Pang, Y., Casal Moura, M., Thompson, G. E., Nelson, D. R., Hummel, A. M., Jenne, D. E., Emerling, D., Volkmuth, W., Robinson, W. H., Specks, U. 2019; 10: 2467

  Abstract

  Mutants of a catalytically inactive variant of Proteinase 3 (PR3)-iPR3-Val103 possessing a Ser195Ala mutation relative to wild-type PR3-Val103-offer insights into how autoantigen PR3 interacts with antineutrophil cytoplasmic antibodies (ANCAs) in granulomatosis with polyangiitis (GPA) and whether such interactions can be interrupted. Here we report that iHm5-Val103, a triple mutant of iPR3-Val103, bound a monoclonal antibody (moANCA518) from a GPA patient on an epitope remote from the mutation sites, whereas the corresponding epitope of iPR3-Val103 was latent to moANCA518. Simulated B-factor analysis revealed that the binding of moANCA518 to iHm5-Val103 was due to increased main-chain flexibility of the latent epitope caused by remote mutations, suggesting rigidification of epitopes with therapeutics to alter pathogenic PR3·ANCA interactions as new GPA treatments.

  View details for DOI 10.3389/fimmu.2019.02467

  View details for PubMedID 31708920

• Autoantibodies to protein-arginine deiminase (PAD) 4 in rheumatoid arthritis: immunological and clinical significance, and potential for precision medicine. Expert review of clinical immunology Martinez-Prat, L. n., Palterer, B. n., Vitiello, G. n., Parronchi, P. n., Robinson, W. H., Mahler, M. n. 2019

  Abstract

  Introduction: The protein-arginine deiminase (PAD) 4 enzyme plays an important role in the pathogenesis of rheumatoid arthritis (RA) and also represents an antigenic target. Anti-PAD4 antibodies can be present in RA and are associated with specific clinical features. Areas covered: This review aims to analyze the current knowledge and recent findings on anti-PAD4 antibodies in RA and their clinical and immunological significance. Expert opinion: Anti-PAD4 antibodies aren't currently used in clinical practice for the management of RA. Nevertheless, there is growing evidence of their relevance in RA, and of their potential utility to improve diagnosis, patient stratification, and prognosis.

  View details for DOI 10.1080/1744666X.2020.1668778

  View details for PubMedID 31524005

• Discovery of Distinct Immune Phenotypes Using Machine Learning in Pulmonary Arterial Hypertension. Circulation research Sweatt, A. J., Hedlin, H. K., Balasubramanian, V. n., Hsi, A. n., Blum, L. K., Robinson, W. H., Haddad, F. n., Hickey, P. M., Condliffe, R. A., Lawrie, A. n., Nicolls, M. R., Rabinovitch, M. n., Khatri, P. n., Zamanian, R. T. 2019

  Abstract

  Accumulating evidence implicates inflammation in pulmonary arterial hypertension (PAH) and therapies targeting immunity are under investigation, though it remains unknown if distinct immune phenotypes exist.Identify PAH immune phenotypes based on unsupervised analysis of blood proteomic profiles.In a prospective observational study of Group 1 PAH patients evaluated at Stanford University (discovery cohort, n=281) and University of Sheffield (validation cohort, n=104) between 2008-2014, we measured a circulating proteomic panel of 48 cytokines, chemokines, and factors using multiplex immunoassay. Unsupervised machine learning (consensus clustering) was applied in both cohorts independently to classify patients into proteomic immune clusters, without guidance from clinical features. To identify central proteins in each cluster, we performed partial correlation network analysis. Clinical characteristics and outcomes were subsequently compared across clusters. Four PAH clusters with distinct proteomic immune profiles were identified in the discovery cohort. Cluster 2 (n=109) had low cytokine levels similar to controls. Other clusters had unique sets of upregulated proteins central to immune networks- cluster 1 (n=58)(TRAIL, CCL5, CCL7, CCL4, MIF), cluster 3 (n=77)(IL-12, IL-17, IL-10, IL-7, VEGF), and cluster 4 (n=37)(IL-8, IL-4, PDGF-β, IL-6, CCL11). Demographics, PAH etiologies, comorbidities, and medications were similar across clusters. Non-invasive and hemodynamic surrogates of clinical risk identified cluster 1 as high-risk and cluster 3 as low-risk groups. Five-year transplant-free survival rates were unfavorable for cluster 1 (47.6%, CI 35.4-64.1%) and favorable for cluster 3 (82.4%, CI 72.0-94.3%)(across-cluster p<0.001). Findings were replicated in the validation cohort, where machine learning classified four immune clusters with comparable proteomic, clinical, and prognostic features.Blood cytokine profiles distinguish PAH immune phenotypes with differing clinical risk that are independent of World Health Organization Group 1 subtypes. These phenotypes could inform mechanistic studies of disease pathobiology and provide a framework to examine patient responses to emerging therapies targeting immunity.

  View details for PubMedID 30661465

• Immunomodulatory receptors are differentially expressed in B and T cell subsets relevant to autoimmune disease. Clinical immunology (Orlando, Fla.) Murphy, K. A., Bhamidiphati, K. n., Rubin, S. J., Kipp, L. n., Robinson, W. H., Lanz, T. V. 2019: 108276

  Abstract

  Inhibitory cell-surface receptors on lymphocytes, often called immune checkpoints, are powerful targets for cancer therapy. Despite their direct involvement in autoimmune pathology, they are currently not exploited therapeutically for autoimmune diseases. Understanding the receptors' expression patterns in health and disease is essential for targeted drug design. Here, we designed three 23-colour flow cytometry panels for peripheral-blood T cells, including 15 lineage-defining markers and 21 immunomodulatory cell-surface receptors, and a 22-marker panel for B cells. Blood samples from healthy individuals, multiple sclerosis (MS), and lupus (SLE) patients were included in the study. Several receptors show differential expression on regulatory T cells (Treg) compared to T helper (Th) 1 and Th17 cells, and functional relevance of this difference could be shown for BTLA and CD5. Unbiased multiparametric analysis revealed a subset of activated CD8+ T cells and a subset of unswitched memory B cells that are diminished in MS and SLE, respectively.

  View details for DOI 10.1016/j.clim.2019.108276

  View details for PubMedID 31669582

• Paraneoplastic and Therapy-Related Immune Complications in Thymic Malignancies. Current treatment options in oncology Lippner, E. A., Lewis, D. B., Robinson, W. H., Katsumoto, T. R. 2019; 20 (7): 62

  Abstract

  The thymus is a key organ involved in establishing central immune tolerance. Thymic epithelial tumors (TETs) include thymomas and thymic carcinomas. Thymomas, which are histologically distinct from thymic carcinomas, lead to dysregulated thymopoiesis via decreased thymic epithelial expression of AIRE and MHC Class II, as well as via alterations in thymic architecture, thereby resulting in autoimmune complications that manifest as paraneoplastic disorders (PNDs). Although progress has been made in elucidating the mechanisms underlying thymoma-associated PNDs, there remains a great need to further define the underlying mechanisms and to identify additional immune biomarkers, such as novel antibodies (in "seronegative" cases) to facilitate diagnosis and monitoring of patients. In addition, a better understanding of the pathogenesis of PNDs could lead to improved treatment strategies for both thymomas and their immune complications. In advanced, refractory cases of TETs (both thymoma and thymic carcinoma), additional therapeutic approaches are needed. Immune checkpoint inhibitors have revolutionized the treatment of several malignancies and hold promise in the treatment of TETs; however, the risks for immune-related adverse events (especially for inducing PNDs as well as in the setting of pre-existing PNDs) underscore the need to optimize patient selection and improve clinical management before there can be widespread acceptance of checkpoint inhibitor therapy in patients with TETs.

  View details for DOI 10.1007/s11864-019-0661-2

  View details for PubMedID 31227926

• Characterizing the BCR repertoire in immune-mediated diseases. Nature reviews. Rheumatology Ramadoss, N. S., Robinson, W. H. 2019

  View details for DOI 10.1038/s41584-019-0339-y

  View details for PubMedID 31780792

• Higher C6 enzyme immunoassay index values correlate with a diagnosis of noncutaneous Lyme disease. Diagnostic microbiology and infectious disease Nigrovic, L. E., Lipsett, S. C., Molins, C. R., Wormser, G. P., Bennett, J. E., Garro, A. C., Levas, M. N., Balamuth, F., Neville, D., Lingampalli, N., Robinson, W. H., Branda, J. A. 2018

  Abstract

  The correlation between the Food and Drug Administration-cleared C6 enzyme immunoassay (EIA) C6 index values and a diagnosis of Lyme disease has not been examined. We used pooled patient-level data from 5 studies of adults and children with Lyme disease and control subjects who were tested with the C6 EIA. We constructed a receiver operating characteristic curve using regression clustered by study and measured the area under the curve (AUC) to examine the accuracy of the C6 index values in differentiating between patients with noncutaneous Lyme disease and control subjects. In the 4821 included patients, the C6 index value had excellent ability to distinguish between patients with noncutaneous Lyme disease and control subjects [AUC 0.99; 95% confidence interval (CI) 0.99-1.00]. An index value cut point of ≥3.0 had a sensitivity of 90.9% (95% CI, 87.8-93.3) and specificity of 99.0% (95% CI, 98.6-99.2%) for Lyme disease.

  View details for DOI 10.1016/j.diagmicrobio.2018.12.001

  View details for PubMedID 30642722

• Affinity Maturation Drives Epitope Spreading and Generation of Proinflammatory Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Elliott, S. E., Kongpachith, S., Lingampalli, N., Adamska, J. Z., Cannon, B. J., Mao, R., Blum, L. K., Robinson, W. H. 2018; 70 (12): 1946–58

  View details for DOI 10.1002/art.40587

  View details for Web of Science ID 000451440200006

• T Cell-Dependent Affinity Maturation and Innate Immune Pathways Differentially Drive Autoreactive B Cell Responses in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Lu, D. R., McDavid, A. N., Kongpachith, S., Lingampalli, N., Glanville, J., Ju, C., Gottardo, R., Robinson, W. H. 2018; 70 (11): 1732–44

  View details for DOI 10.1002/art.40578

  View details for Web of Science ID 000448797600005

• Direct Diagnostic Tests for Lyme Disease. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America Schutzer, S. E., Body, B. A., Boyle, J., Branson, B. M., Dattwyler, R. J., Fikrig, E., Gerald, N. J., Gomes-Solecki, M., Kintrup, M., Ledizet, M., Levin, A. E., Lewinski, M., Liotta, L. A., Marques, A., Mead, P. S., Mongodin, E. F., Pillai, S., Rao, P., Robinson, W. H., Roth, K. M., Schriefer, M. E., Slezak, T., Snyder, J. L., Steere, A. C., Witkowski, J., Wong, S. J., Branda, J. A. 2018

  Abstract

  Borrelia burgdorferi was discovered to be the cause of Lyme disease in 1983, leading to seroassays. The 1994 serodiagnostic testing guidelines predated a full understanding of key B. burgdorferi antigens and have a number of shortcomings. These serologic tests cannot distinguish active infection, past infection, or reinfection. Reliable direct-detection methods for active B. burgdorferi infection have been lacking in the past but are needed and appear achievable. New approaches have effectively been applied to other emerging infections and show promise in direct detection of B. burgdorferi infections.

  View details for PubMedID 30307486

• Combination of anti-citrullinated protein antibodies and rheumatoid factor is associated with increased systemic inflammatory mediators and more updates rapid progression from preclinical to clinical rheumatoid arthritis CLINICAL IMMUNOLOGY Lingampalli, N., Sokolove, J., Lahey, L. J., Edison, J. D., Gilliland, W. R., Holers, V., Deane, K. D., Robinson, W. H. 2018; 195: 119–26

  View details for DOI 10.1016/j.clim.2018.05.004

  View details for Web of Science ID 000444665600016

• Engineered DNA plasmid reduces immunity to dystrophin while improving muscle force in a model of gene therapy of Duchenne dystrophy PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Ho, P. P., Lahey, L. J., Mourkioti, F., Kraft, P. E., Filareto, A., Brandt, M., Magnusson, K. G., Finn, E. E., Chamberlain, J. S., Robinson, W. H., Blau, H. M., Steinman, L. 2018; 115 (39): E9182–E9191

  View details for DOI 10.1073/pnas.1808648115

  View details for Web of Science ID 000445545200019

• Engineered DNA plasmid reduces immunity to dystrophin while improving muscle force in a model of gene therapy of Duchenne dystrophy. Proceedings of the National Academy of Sciences of the United States of America Ho, P. P., Lahey, L. J., Mourkioti, F., Kraft, P. E., Filareto, A., Brandt, M., Magnusson, K. E., Finn, E. E., Chamberlain, J. S., Robinson, W. H., Blau, H. M., Steinman, L. 2018

  Abstract

  In gene therapy for Duchenne muscular dystrophy there are two potential immunological obstacles. An individual with Duchenne muscular dystrophy has a genetic mutation in dystrophin, and therefore the wild-type protein is "foreign," and thus potentially immunogenic. The adeno-associated virus serotype-6 (AAV6) vector for delivery of dystrophin is a viral-derived vector with its own inherent immunogenicity. We have developed a technology where an engineered plasmid DNA is delivered to reduce autoimmunity. We have taken this approach into humans, tolerizing to myelin proteins in multiple sclerosis and to proinsulin in type 1 diabetes. Here, we extend this technology to a model of gene therapy to reduce the immunogenicity of the AAV vector and of the wild-type protein product that is missing in the genetic disease. Following gene therapy with systemic administration of recombinant AAV6-microdystrophin to mdx/mTRG2 mice, we demonstrated the development of antibodies targeting dystrophin and AAV6 capsid in control mice. Treatment with the engineered DNA construct encoding microdystrophin markedly reduced antibody responses to dystrophin and to AAV6. Muscle force in the treated mice was also improved compared with control mice. These data highlight the potential benefits of administration of an engineered DNA plasmid encoding the delivered protein to overcome critical barriers in gene therapy to achieve optimal functional gene expression.

  View details for PubMedID 30181272

• Rheumatoid arthritis and the mucosal origins hypothesis: protection turns to destruction NATURE REVIEWS RHEUMATOLOGY Holers, V., Demoruelle, M., Kuhn, K. A., Buckner, J. H., Robinson, W. H., Okamoto, Y., Norris, J. M., Deane, K. D. 2018; 14 (9): 542–57

  Abstract

  Individuals at high risk of developing seropositive rheumatoid arthritis (RA) can be identified for translational research and disease prevention studies through the presence of highly informative and predictive patterns of RA-related autoantibodies, especially anti-citrullinated protein antibodies (ACPAs), in the serum. In serologically positive individuals without arthritis, designated ACPA positive at risk, the presence of mucosal inflammatory processes associated with the presence of local ACPA production has been demonstrated. In other at-risk populations, local RA-related autoantibody production is present even in the absence of serum autoantibodies. Additionally, a proportion of at-risk individuals exhibit local mucosal ACPA production in the lung, as well as radiographic small-airway disease, sputum hypercellularity and increased neutrophil extracellular trap formation. Other mucosal sites in at-risk individuals also exhibit autoantibody production, inflammation and/or evidence of dysbiosis. As the proportion of individuals who exhibit such localized inflammation-associated ACPA production is substantially higher than the likelihood of an individual developing future RA, this finding raises the hypothesis that mucosal ACPAs have biologically relevant protective roles. Identifying the mechanisms that drive both the generation and loss of externally focused mucosal ACPA production and promote systemic autoantibody expression and ultimately arthritis development should provide insights into new therapeutic approaches to prevent RA.

  View details for PubMedID 30111803

• Characterization of Monoclonal Anti-PAD4 Autoantibodies from Rheumatoid Arthritis Patients: Functional Implications for Citrullination and Disease Progression Gomez, A., Kongpachith, S., Lingampalli, N., Cisar, C., Robinson, W. H. WILEY. 2018

  View details for Web of Science ID 000447268905113

• Expanded Autoantibody Profiles Predict Treatment Response to Biologic Therapy in Rheumatoid Arthritis Mikuls, T. R., Bath, A., England, B. R., Duryee, M. J., Hunter, C. D., Kremer, J., Pappas, D. A., Robinson, W. H., Curtis, J. R., Thiele, G. M. WILEY. 2018

  View details for Web of Science ID 000447268901110

• Histologic and Transcriptional Evidence of Subclinical Synovial Inflammation in Rheumatoid Arthritis Patients in Clinical Remission Orange, D., Agius, P., DiCarlo, E. F., Mirza, S. Z., Rozo, C., Pannellini, T., Figgie, M. P., Robinson, W. H., Szymonifka, J., Bykerk, V. P., Donlin, L. T., Goodman, S. M. WILEY. 2018

  View details for Web of Science ID 000447268903133

• Antibody Repertoire Dynamics in Systemic Sclerosis after Myeloablative Autologous Hematopoietic Stem-Cell Transplantation or Cyclophosphamide Treatment Adamska, J. Z., Crofford, L., Furst, D. E., Goldmuntz, E., Keyes-Elstein, L., Mayes, M. D., McSweeney, P., Nash, R., Pinckney, A., Welch, B., Sullivan, K., Robinson, W. H. WILEY. 2018

  View details for Web of Science ID 000447268902209

• Serum Cytokine and Chemokine Concentrations Predict Incident Cancer in US Veterans with Rheumatoid Arthritis England, B. R., Sayles, H., Roul, P., Ganti, A., Sokolove, J., Robinson, W. H., Cannon, G. W., Sauer, B., Baker, J. F., Thiele, G. M., Mikuls, T. R. WILEY. 2018

  View details for Web of Science ID 000447268901413

• Characterization of Preferential Recognition of a Chimeric Recombinant Proteinase 3 Variant By Anti-Neutrophil Cytoplasmic Antibodies Thompson, G., Moura, M., Nelson, D., Hummel, A., Jenne, D. E., Fervenza, F., Hoffman, G. S., Kallenberg, C. M., Langford, C., McCune, J. W., Merkel, P. A., Monach, P. A., Seo, P., Spiera, R. F., St Clair, E., Ytterberg, S. R., Stone, J. H., Robinson, W. H., Pang, Y., Specks, U. WILEY. 2018

  View details for Web of Science ID 000447268902008

• Vertebral Compression Fractures and Antibodies to Citrullinated Vimentin in Established Rheumatoid Arthritis Cheah, J., Carrino, J. A., Robinson, W. H., Orange, D., Szymonifka, J., Stein, E. M., Goodman, S. M. WILEY. 2018

  View details for Web of Science ID 000447268904070

• Rheumatoid Factor Peptide Expression By Quantitative Proteomics Delineates Responsive and Resistant Clones after Treatment of Early Rheumatoid Arthritis Wang, J., Murray-Brown, W., Colella, A., Smith, M. D., Chataway, T., Walker, J., Robinson, W. H., Wechalekar, M. D., Gordon, T. WILEY. 2018

  View details for Web of Science ID 000447268903254

• Antibodies to Citrullinated Protein Antigens Are Associated with Risk of Cardiovascular Disease in Community-Dwelling Women: The Multi-Ethnic Study of Atherosclerosis Hughes-Austin, J. M., Katz, R., Firestein, G. S., Criqui, M. H., Robinson, W. H., Ix, J. H. WILEY. 2018

  View details for Web of Science ID 000447268900209

• Plasmablast antibody repertoires in elderly influenza vaccine responders exhibit restricted diversity but increased breadth of binding across influenza strains CLINICAL IMMUNOLOGY Ju, C., Blum, L. K., Kongpachith, S., Lingampalli, N., Mao, R., Brodin, P., Dekker, C. L., Davis, M. M., Robinson, W. H. 2018; 193: 70–79

  View details for DOI 10.1016/j.clim.2018.01.011

  View details for Web of Science ID 000440126900010

• Robust B Cell Responses Predict Rapid Resolution of Lyme Disease FRONTIERS IN IMMUNOLOGY Blum, L. K., Adamska, J. Z., Martin, D. S., Rebman, A. W., Elliott, S. E., Cao, R. L., Embers, M. E., Aucott, J. N., Soloski, M. J., Robinson, W. H. 2018; 9

  View details for DOI 10.3389/fimmu.2018.01634

  View details for Web of Science ID 000439051400001

• Robust B Cell Responses Predict Rapid Resolution of Lyme Disease. Frontiers in immunology Blum, L. K., Adamska, J. Z., Martin, D. S., Rebman, A. W., Elliott, S. E., Cao, R. R., Embers, M. E., Aucott, J. N., Soloski, M. J., Robinson, W. H. 2018; 9: 1634

  Abstract

  Lyme disease (Borrelia burgdorferi infection) is increasingly recognized as a significant source of morbidity worldwide. Here, we show that blood plasmablasts and CD27- memory B cells are elevated in untreated Lyme disease, with higher plasmablast levels associated with more rapid resolution of clinical symptoms. Stronger serum reactivity to surface proteins and peptides from B. burgdorferi was also associated with faster resolution of clinical symptoms. Through molecular identifier-enabled antibody heavy-chain sequencing of bulk B cells and single-cell paired-chain antibody sequencing of blood plasmablasts, we characterized immunoglobulin gene usage patterns specific to B. burgdorferi infection. Recombinantly expressed antibodies from expanded lineages bound B. burgdorferi antigens, confirming that these clones are driven by the infection. Furthermore, recombinant sequence-derived antibodies were functional, inhibiting growth of B. burgdorferi in vitro. Elevations and clonal expansion of blood plasmablasts were associated with rapid return to health, while poor plasmablast responses were associated with a longer duration of symptoms following treatment. Plasmablasts induced by B. burgdorferi infection showed preferential antibody gene segment usage, while bulk sequencing of total B cells revealed convergent CDR3 motifs specific to B. burgdorferi-infected patients. Our results show that robust plasmablast responses encoding Bb-static antibodies are associated with more rapid resolution of Lyme disease, and these antibodies could provide the basis for next-generation therapeutics for Lyme disease.

  View details for DOI 10.3389/fimmu.2018.01634

  View details for PubMedID 30072990

  View details for PubMedCentralID PMC6060717

• Methods for high-dimensonal analysis of cells dissociated from cyropreserved synovial tissue ARTHRITIS RESEARCH & THERAPY Donlin, L. T., Rao, D. A., Wei, K., Slowikowski, K., McGeachy, M. J., Turner, J. D., Meednu, N., Mizoguchi, F., Gutierrez-Arcelus, M., Lieb, D. J., Keegan, J., Muskat, K., Hillman, J., Rozo, C., Ricker, E., Eisenhaure, T. M., Li, S., Browne, E. P., Chicoine, A., Sutherby, D., Noma, A., Nusbaum, C., Kelly, S., Pernis, A. B., Ivashkiv, L. B., Goodman, S. M., Robinson, W. H., Utz, P. J., Lederer, J. A., Gravallese, E. M., Boyce, B. F., Hacohen, N., Pitzalis, C., Gregersen, P. K., Firestein, G. S., Raychaudhuri, S., Moreland, L. W., Holers, V., Bykerk, V. P., Filer, A., Boyle, D. L., Brenner, M. B., Anolik, J. H., Accelerating Med Partnership RA SL 2018; 20: 139

  Abstract

  Detailed molecular analyses of cells from rheumatoid arthritis (RA) synovium hold promise in identifying cellular phenotypes that drive tissue pathology and joint damage. The Accelerating Medicines Partnership RA/SLE Network aims to deconstruct autoimmune pathology by examining cells within target tissues through multiple high-dimensional assays. Robust standardized protocols need to be developed before cellular phenotypes at a single cell level can be effectively compared across patient samples.Multiple clinical sites collected cryopreserved synovial tissue fragments from arthroplasty and synovial biopsy in a 10% DMSO solution. Mechanical and enzymatic dissociation parameters were optimized for viable cell extraction and surface protein preservation for cell sorting and mass cytometry, as well as for reproducibility in RNA sequencing (RNA-seq). Cryopreserved synovial samples were collectively analyzed at a central processing site by a custom-designed and validated 35-marker mass cytometry panel. In parallel, each sample was flow sorted into fibroblast, T-cell, B-cell, and macrophage suspensions for bulk population RNA-seq and plate-based single-cell CEL-Seq2 RNA-seq.Upon dissociation, cryopreserved synovial tissue fragments yielded a high frequency of viable cells, comparable to samples undergoing immediate processing. Optimization of synovial tissue dissociation across six clinical collection sites with ~ 30 arthroplasty and ~ 20 biopsy samples yielded a consensus digestion protocol using 100 μg/ml of Liberase™ TL enzyme preparation. This protocol yielded immune and stromal cell lineages with preserved surface markers and minimized variability across replicate RNA-seq transcriptomes. Mass cytometry analysis of cells from cryopreserved synovium distinguished diverse fibroblast phenotypes, distinct populations of memory B cells and antibody-secreting cells, and multiple CD4+ and CD8+ T-cell activation states. Bulk RNA-seq of sorted cell populations demonstrated robust separation of synovial lymphocytes, fibroblasts, and macrophages. Single-cell RNA-seq produced transcriptomes of over 1000 genes/cell, including transcripts encoding characteristic lineage markers identified.We have established a robust protocol to acquire viable cells from cryopreserved synovial tissue with intact transcriptomes and cell surface phenotypes. A centralized pipeline to generate multiple high-dimensional analyses of synovial tissue samples collected across a collaborative network was developed. Integrated analysis of such datasets from large patient cohorts may help define molecular heterogeneity within RA pathology and identify new therapeutic targets and biomarkers.

  View details for PubMedID 29996944

• Chemerin 156F, generated by chymase cleavage of prochemerin, is elevated in joint fluids of arthritis patients. Arthritis research & therapy Zhao, L., Yamaguchi, Y., Ge, X., Robinson, W. H., Morser, J., Leung, L. L. 2018; 20 (1): 132

  Abstract

  BACKGROUND: Chemerin is a chemoattractant involved in immunity that also functions as an adipokine. Chemerin is secreted as an inactive precursor (chem163S), and its activation requires proteolytic cleavages at its C-terminus, involving proteases in coagulation, fibrinolysis, and inflammation. Previously, we found chem158K was the dominant chemerin form in synovial fluids from patients with arthritis. In this study, we aimed to characterize a distinct cleaved chemerin form, chem156F, in osteoarthritis (OA) and rheumatoid arthritis (RA).METHODS: Purified chem156F was produced in transfected CHO cells. To quantify chem156F in OA and RA samples, we developed a specific ELISA for chem156F using antibody raised against a peptide representing the C-terminus of chem156F.RESULTS: Ca2+ mobilization assays showed that the EC50 values for chem163S, chem156F, and chem157S were 252±141nM, 133±41.5nM, and 5.83±2.48nM, respectively. chem156F was more active than its precursor, chem163S, but very much less potent than chem157S, the most active chemerin form. Chymase was shown to be capable of cleaving chem163S at a relevant rate. Using the chem156F ELISA we found a substantial amount of chem156F present in synovial fluids from patients with OA and RA, 24.06±5.51ng/ml and 20.35±5.19ng/ml (mean±SEM, n=25) respectively, representing 20% of total chemerin in OA and 76.7% of chemerin in RA synovial fluids.CONCLUSIONS: Our data show that chymase cleavage of chem163S to partially active chem156F can be found in synovial fluids where it can play a role in modulation of the inflammation in joints.

  View details for PubMedID 29973268

• Chemerin 156F, generated by chymase cleavage of prochemerin, is elevated in joint fluids of arthritis patients ARTHRITIS RESEARCH & THERAPY Zhao, L., Yamaguchi, Y., Ge, X., Robinson, W. H., Morser, J., Leung, L. K. 2018; 20

  View details for DOI 10.1186/s13075-018-1615-y

  View details for Web of Science ID 000437485000001

• Increased somatic hypermutation in the immunoglobulin sequences of melanoma patients who have durable response to checkpoint inhibitor therapy Ngan Nguyen, Mamchak, A., Severgnini, M., Williamson, K. S., Wu, X., Drabek, E. F., Manos, M., Chen, X., Tang, X., Amiri, Z., Carroll, C., Leung, Y., Kim, D., Volkmuth, W., Greenberg, N., Emerling, D., Robinson, W. H., Cavet, G., Hodi, F. AMER ASSOC CANCER RESEARCH. 2018

  View details for DOI 10.1158/1538-7445.AM2018-615

  View details for Web of Science ID 000468818902128

• Mining the cancer immuno-responsome: The identification of functional antitumor antibodies from patients receiving checkpoint inhibitors Baia, G., Manning-Bog, A., Scholz, A., DeFalco, J., Harbell, M., Zhang, D., Chu, F., Millare, B., Sumi, M., Zuno, P., Sapugay, J., Kim, D., Leung, Y., Jiang, S., Tang, X., Williamson, K., Chen, X., Carroll, S., Dowd, C., Dhawan, I., Benjamin, J., Santo, G., Haaser, N., Ngan Nguyen, Giladi, E., Minor, D., Tan, Y., Sokolove, J. B., Steinman, L., Serafini, T., Cavet, G., Greenberg, N. M., Glanville, J., Volkmuth, W., Emerling, D. E., Robinson, W. H. AMER ASSOC CANCER RESEARCH. 2018

  View details for DOI 10.1158/1538-7445.AM2018-3966

  View details for Web of Science ID 000468819502221

• Plasma adiponectin levels are associated with circulating inflammatory cytokines in autoantibody positive first-degree relatives of rheumatoid arthritis patients PLOS ONE Hughes-Austin, J. M., Deane, K. D., Giles, J. T., Derber, L. A., Zerbe, G. O., Dabelea, D. M., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M. 2018; 13 (6): e0199578

  Abstract

  Extra-articular manifestations of rheumatoid arthritis (RA), potentially due to systemic inflammation, include cardiovascular disease and sarcopenic obesity. Adiponectin, an adipose-derived cytokine, has been implicated in inflammatory processes in RA, but little is known regarding its association with inflammation in a pre-clinical period. Therefore, we investigated whether adiponectin was associated with inflammatory markers in individuals at risk for RA, and whether RA-related autoimmunity modifies these associations.We analyzed samples from 144 first-degree relatives (FDRs) of RA probands, of whom 23 were positive for anti-cyclic citrullinated peptide antibody and/or ≥ 2 rheumatoid factor isotypes (IgM, IgG or IgA). We called this phenotype the 'high risk autoantibody profile (HRP)' as it has been shown in prior work to be >96% specific for future RA. We measured adiponectin, cytokines, and high-sensitivity C-reactive protein (hsCRP). Using linear mixed effects models, we evaluated interaction between HRP positivity and adiponectin on inflammatory markers, adjusting for age, sex, ethnicity, body mass index, pack-years smoking, and use of cholesterol-lowering medications.In everyone, adiponectin concentration was inversely associated with hsCRP and IL-1β in adjusted models, where a 1% higher adiponectin was associated with a 26% lower hsCRP (p = 0.04) and a 26% lower IL-1β (p = 0.04). Significant interactions between HRP and adiponectin for associations with GM-CSF, IL-6, and IL-9 were detected in fully adjusted models (p = 0.0006, p = 0.006, p = 0.01, respectively). In HRP positive FDRs but not HRP negative FDRs, a 1% higher adiponectin was associated with 97% higher GM-CSF, 73% higher IL-6, and 54% higher IL-9 concentrations.Adiponectin associates with inflammatory markers, and these associations differ in individuals with a high-risk autoantibody profile compared with those without. The interaction between adiponectin and autoimmunity warrants further investigation into the potential systemic effects of RA-related autoantibodies and adiponectin on inflammation in the absence of clinically apparent RA.

  View details for PubMedID 29940013

• Combination of anti-citrullinated protein antibodies and rheumatoid factor is associated with increased systemic inflammatory mediators and more rapid progression from preclinical to clinical rheumatoid arthritis. Clinical immunology (Orlando, Fla.) Lingampalli, N., Sokolove, J., Lahey, L. J., Edison, J. D., Gilliland, W. R., Michael Holers, V., Deane, K. D., Robinson, W. H. 2018

  Abstract

  The development of rheumatoid factor (RF) and/or anti-citrullinated protein antibodies (ACPAs) can be observed years prior to clinical diagnosis of rheumatoid arthritis (RA). Nevertheless, the interaction between these two autoantibodies and their combined effect on development of RA is unclear. We measured RF, cytokines, and ACPA subtypes in serial pre-clinical serum samples collected from 83 US veterans who all developed RA. Levels of cytokines and ACPAs were compared between the following groups: anti-cyclic citrullinated peptide (anti-CCP)-/RF- (double negative), anti-CCP+/RF-, anti-CCP-/RF+, or anti-CCP+/RF+ (double-positive). The double-positive subgroup had significantly higher levels of 20 inflammatory cytokines and 29 ACPA reactivities, and the shortest interval, 1.3 years, between the preclinical sample timepoint and diagnosis of RA. Thus, the combined presence of ACPAs and RF is associated with a more rapid progression to RA, suggesting that anti-CCP+/RF+ individuals have a more advanced preclinical disease state and that the onset of RA may be imminent.

  View details for PubMedID 29842946

• Impact on immune effectors of anti-tumor antibodies isolated from cancer patients. Manning-Bog, A., Baia, G., Defalco, J., Aydin, I. T., Harbell, M. G., Zuno-Mitchell, P., Sapugay, J., Zhang, D., Chu, F., Millare, B., Scholz, A., Sumi, M., Fernandez, S., Haaser, N., Benjamin, J., Nguyen, N., Volkmuth, W., Robinson, W. H., Greenberg, N. M., Emerling, D. AMER SOC CLINICAL ONCOLOGY. 2018

  View details for DOI 10.1200/JCO.2018.36.15_suppl.e15015

  View details for Web of Science ID 000442916004639

• Revealing the specificity of regulatory T cells in murine autoimmune diabetes PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Spence, A., Purtha, W., Tam, J., Dong, S., Kim, Y., Ju, C., Sterling, T., Nakayama, M., Robinson, W. H., Bluestone, J. A., Anderson, M. S., Tang, Q. 2018; 115 (20): 5265–70

  Abstract

  Regulatory T cells (Tregs) control organ-specific autoimmunity in a tissue antigen-specific manner, yet little is known about their specificity in a natural repertoire. In this study, we used the nonobese diabetic (NOD) mouse model of autoimmune diabetes to investigate the antigen specificity of Tregs present in the inflamed tissue, the islets of Langerhans. Compared with Tregs present in spleen and lymph node, Tregs in the islets showed evidence of antigen stimulation that correlated with higher proliferation and expression of activation markers CD103, ICOS, and TIGIT. T cell receptor (TCR) repertoire profiling demonstrated that islet Treg clonotypes are expanded in the islets, suggesting localized antigen-driven expansion in inflamed islets. To determine their specificity, we captured TCRαβ pairs from islet Tregs using single-cell TCR sequencing and found direct evidence that some of these TCRs were specific for islet-derived antigens including insulin B:9-23 and proinsulin. Consistently, insulin B:9-23 tetramers readily detected insulin-specific Tregs in the islets of NOD mice. Lastly, islet Tregs from prediabetic NOD mice were effective at preventing diabetes in Treg-deficient NOD.CD28-/- recipients. These results provide a glimpse into the specificities of Tregs in a natural repertoire that are crucial for opposing the progression of autoimmune diabetes.

  View details for PubMedID 29712852

• Identification of Three Rheumatoid Arthritis Disease Subtypes by Machine Learning Integration of Synovial Histologic Features and RNA Sequencing Data ARTHRITIS & RHEUMATOLOGY Orange, D. E., Agius, P., DiCarlo, E. F., Robine, N., Geiger, H., Szymonifka, J., McNamara, M., Cummings, R., Andersen, K. M., Mirza, S., Figgie, M., Ivashkiv, L. B., Pernis, A. B., Jiang, C. S., Frank, M. O., Darnell, R. B., Lingampali, N., Robinson, W. H., Gravallese, E., Bykerk, V. P., Goodman, S. M., Donlin, L. T., Accelerating Medicines Partnership 2018; 70 (5): 690–701

  Abstract

  In this study, we sought to refine histologic scoring of rheumatoid arthritis (RA) synovial tissue by training with gene expression data and machine learning.Twenty histologic features were assessed in 129 synovial tissue samples (n = 123 RA patients and n = 6 osteoarthritis [OA] patients). Consensus clustering was performed on gene expression data from a subset of 45 synovial samples. Support vector machine learning was used to predict gene expression subtypes, using histologic data as the input. Corresponding clinical data were compared across subtypes.Consensus clustering of gene expression data revealed 3 distinct synovial subtypes, including a high inflammatory subtype characterized by extensive infiltration of leukocytes, a low inflammatory subtype characterized by enrichment in pathways including transforming growth factor β, glycoproteins, and neuronal genes, and a mixed subtype. Machine learning applied to histologic features, with gene expression subtypes serving as labels, generated an algorithm for the scoring of histologic features. Patients with the high inflammatory synovial subtype exhibited higher levels of markers of systemic inflammation and autoantibodies. C-reactive protein (CRP) levels were significantly correlated with the severity of pain in the high inflammatory subgroup but not in the others.Gene expression analysis of RA and OA synovial tissue revealed 3 distinct synovial subtypes. These labels were used to generate a histologic scoring algorithm in which the histologic scores were found to be associated with parameters of systemic inflammation, including the erythrocyte sedimentation rate, CRP level, and autoantibody levels. Comparison of gene expression patterns to clinical features revealed a potentially clinically important distinction: mechanisms of pain may differ in patients with different synovial subtypes.

  View details for PubMedID 29468833

• Circulating plasmablasts are elevated and produce pathogenic anti-endothelial cell autoantibodies in idiopathic pulmonary arterial hypertension EUROPEAN JOURNAL OF IMMUNOLOGY Blum, L. K., Cao, R. L., Sweatt, A. J., Bill, M., Lahey, L. J., Hsi, A. C., Lee, C. S., Kongpachith, S., Ju, C., Mao, R., Wong, H. H., Nicolls, M. R., Zamanian, R. T., Robinson, W. H. 2018; 48 (5): 874–84

  View details for DOI 10.1002/eji.201747460

  View details for Web of Science ID 000435725100016

• Advances in Serodiagnostic Testing for Lyme Disease Are at Hand CLINICAL INFECTIOUS DISEASES Branda, J. A., Body, B. A., Boyle, J., Branson, B. M., Dattwyler, R. J., Fikrig, E., Gerald, N. J., Gomes-Solecki, M., Kintrup, M., Ledizet, M., Levin, A. E., Lewinski, M., Liotta, L. A., Marques, A., Mead, P. S., Mongodin, E. F., Pillai, S., Rao, P., Robinson, W. H., Roth, K. M., Schriefer, M. E., Slezak, T., Snyder, J., Steere, A. C., Witkowski, J., Wong, S. J., Schutzer, S. E. 2018; 66 (7): 1133–39

  Abstract

  The cause of Lyme disease, Borrelia burgdorferi, was discovered in 1983. A 2-tiered testing protocol was established for serodiagnosis in 1994, involving an enzyme immunoassay (EIA) or indirect fluorescence antibody, followed (if reactive) by immunoglobulin M and immunoglobulin G Western immunoblots. These assays were prepared from whole-cell cultured B. burgdorferi, lacking key in vivo expressed antigens and expressing antigens that can bind non-Borrelia antibodies. Additional drawbacks, particular to the Western immunoblot component, include low sensitivity in early infection, technical complexity, and subjective interpretation when scored by visual examination. Nevertheless, 2-tiered testing with immunoblotting remains the benchmark for evaluation of new methods or approaches. Next-generation serologic assays, prepared with recombinant proteins or synthetic peptides, and alternative testing protocols, can now overcome or circumvent many of these past drawbacks. This article describes next-generation serodiagnostic testing for Lyme disease, focusing on methods that are currently available or near-at-hand.

  View details for PubMedID 29228208

• Antibody Responses to Citrullinated and Noncitrullinated Antigens in the Sputum of Subjects With Rheumatoid Arthritis and Subjects at Risk for Development of Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Demoruelle, M., Bowers, E., Lahey, L. J., Sokolove, J., Purmalek, M., Seto, N. L., Weisman, M. H., Norris, J. M., Kaplan, M. J., Holers, V., Robinson, W. H., Deane, K. D. 2018; 70 (4): 516–27

  Abstract

  The location and mechanisms involved in the initial generation of autoantibodies to citrullinated and noncitrullinated proteins/peptides during the natural history of rheumatoid arthritis (RA) development is incompletely understood. This study sought to explore individual antibody responses to citrullinated and noncitrullinated proteins/peptides in the sputum and associations with neutrophil extracellular traps (NETs) in subjects at risk for the future development of RA.Serum and sputum samples were obtained from 41 RA-free subjects who were considered at risk for the development of RA based on familial or serologic risk factors, from 20 subjects classified as having RA, and from 22 healthy control subjects. Samples were evaluated using a bead-based array for IgG reactivity to 29 citrullinated proteins/peptides and 21 noncitrullinated proteins/peptides. Cutoff levels for antibody positivity were established in a separate control group. NET levels in the sputum were measured using sandwich enzyme-linked immunosorbent assays that quantitate DNA-myeloperoxidase and DNA-neutrophil elastase complexes.In at-risk subjects, antibody responses to the citrullinated forms of fibrinogen, apolipoprotein E, and fibronectin were highly prevalent. The most citrulline-specific antibodies in the sputum of at-risk subjects were those to fibrinogen, vimentin, and peptides of fibrinogen A and apolipoprotein A1. Patterns of sputum autoantibody positivity differed between at-risk subjects and subjects with RA. In at-risk subjects, increasing sputum NET levels significantly correlated with several citrullinated and some noncitrullinated antibody reactivities.These findings suggest that sputum antibody reactivity to particular citrullinated and noncitrullinated proteins/peptides is specific for RA and for subjects at risk of RA, and the association of these proteins/peptides with NETs may be a key feature of early RA-related autoimmunity in the lung. These results further support the hypothesis that the lung plays a role in early RA-related autoimmunity.

  View details for PubMedID 29266801

  View details for PubMedCentralID PMC5876113

• Response to comment on "Synovial fibroblast-neutrophil interactions promote pathogenic adaptive immunity in rheumatoid arthritis" SCIENCE IMMUNOLOGY Carmona-Rivera, C., Bicker, K. L., Thompson, P. R., Buckner, J. H., Robinson, W. H., Fox, D. A., Kaplan, M. J. 2018; 3 (21)

  Abstract

  The citrullinome cargo in neutrophil extracellular traps varies according to disease condition and stimulation conditions.

  View details for PubMedID 29602804

• Plasmablast antibody repertoires in elderly influenza vaccine responders exhibit restricted diversity but increased breadth of binding across influenza strains. Clinical immunology (Orlando, Fla.) Ju, C., Blum, L. K., Kongpachith, S., Lingampalli, N., Mao, R., Brodin, P., Dekker, C. L., Davis, M. M., Robinson, W. H. 2018

  Abstract

  Seasonal influenza vaccines elicit antibody responses that can prevent infection, but their efficacy is reduced in the elderly. While a subset of elderly individuals can still mount sufficient vaccine-induced antibody responses, little is known about the properties of the vaccine-induced antibody repertoires in elderly as compared to young responders. To gain insights into the effects of aging on influenza vaccine-induced antibody responses, we used flow cytometry and a cell-barcoding method to sequence antibody heavy and light chain gene pairs expressed by individual blood plasmablasts generated in response to influenza vaccination in elderly (aged 70-89) and young (aged 20-29) responders. We found similar blood plasmablast levels in the elderly and young responders seven days post vaccination. Informatics analysis revealed increased clonality, but similar heavy chain V(D)J gene usage in the elderly as compared to young vaccine responders. Although the elderly responders exhibited decreased antibody sequence diversity and fewer consequential mutations relative to young responders, recombinant antibodies from elderly responders bound a broader range of influenza strain HAs. Thus elderly influenza vaccine responders mount plasmablast responses with restricted diversity but with an increased breadth of binding across influenza strains. Our results suggest that the ability to generate plasmablast responses encoding cross-strain binding antibodies likely represents a mechanism important to vaccine responses in the elderly.

  View details for PubMedID 29410330

• Men and Women Differ in the Biochemical Composition of Platelet-Rich Plasma AMERICAN JOURNAL OF SPORTS MEDICINE Xiong, G., Lingampalli, N., Koltsov, J. B., Leung, L. L., Bhutani, N., Robinson, W. H., Chu, C. R. 2018; 46 (2): 409–19

  Abstract

  Autologous platelet-rich plasma (PRP) is widely used for a variety of clinical applications. However, clinical outcome studies have not consistently shown positive effects. The composition of PRP differs based on many factors. An improved understanding of factors influencing the composition of PRP is important for the optimization of PRP use.Age and sex influence the PRP composition in healthy patients.Controlled laboratory study.Blood from 39 healthy patients was collected at a standardized time and processed into leukocyte-poor PRP within 1 hour of collection using the same laboratory centrifuge protocol and frozen for later analysis. Eleven female and 10 male patients were "young" (aged 18-30 years), while 8 male and 10 female patients were "older" (aged 45-60 years). Thawed PRP samples were assessed for cytokine and growth factor levels using a multiplex assay and enzyme-linked immunosorbent assay. The platelet count and high-sensitivity C-reactive protein levels were measured. Two-way analysis of variance determined age- and sex-based differences.Platelet and high-sensitivity C-reactive protein concentrations were similar in PRP between the groups ( P = .234). Male patients had higher cytokine and growth factor levels in PRP compared with female patients for inflammatory cytokines such as interleukin-1 beta (IL-1β) (9.83 vs 7.71 pg/mL, respectively; P = .008) and tumor necrosis factor-alpha (TNF-α) (131.6 vs 110.5 pg/mL, respectively; P = .048); the anti-inflammatory IL-1 receptor antagonist protein (IRAP) (298.0 vs 218.0 pg/mL, respectively; P < .001); and growth factors such as fibroblast growth factor-basic (FGF-basic) (237.9 vs 194.0 pg/mL, respectively; P = .01), platelet-derived growth factor (PDGF-BB) (3296.2 vs 2579.3 pg/mL, respectively; P = .087), and transforming growth factor-beta 1 (TGF-β1) (118.8 vs 92.8 ng/mL, respectively; P = .002). Age- but not sex-related differences were observed for insulin-like growth factor-1 (IGF-1) ( P < .001). Age and sex interaction terms were not significant. While mean differences were significant, there was also substantial intragroup variability.This study in healthy patients shows differences in the composition of PRP between men and women, with sex being a greater factor than age. There was also proteomic variability within the groups. These data support a personalized approach to PRP treatment and highlight the need for a greater understanding of the relationships between proteomic factors in PRP and clinical outcomes.Variability in the proteomic profile of PRP may affect tissue and clinical responses to treatment. These data suggest that clinical studies should account for the composition of PRP used.

  View details for PubMedID 29211968

• Identification of differentially expressed micro-RNA in rotator cuff tendinopathy MLTJ-MUSCLES LIGAMENTS AND TENDONS JOURNAL Hall, K. E., Sarkissian, E. J., Sharpe, O., Robinson, W. H., Abrams, G. D. 2018; 8 (1): 8–14

  View details for DOI 10.11138/mltj/2018.8.1.008

  View details for Web of Science ID 000431248200002

• Circulating plasmablasts are elevated and produce pathogenic anti-endothelial cell autoantibodies in idiopathic pulmonary arterial hypertension. European journal of immunology Blum, L. K., Cao, R. R., Sweatt, A. J., Bill, M. n., Lahey, L. J., Hsi, A. C., Lee, C. S., Kongpachith, S. n., Ju, C. H., Mao, R. n., Wong, H. H., Nicolls, M. R., Zamanian, R. T., Robinson, W. H. 2018

  Abstract

  Idiopathic pulmonary arterial hypertension (IPAH) is a devastating pulmonary vascular disease in which autoimmune and inflammatory phenomena are implicated. B cells and autoantibodies have been associated with IPAH and identified as potential therapeutic targets. However, the specific populations of B cells involved and their roles in disease pathogenesis are not clearly defined. We aimed to assess the levels of activated B cells (plasmablasts) in IPAH, and to characterize recombinant antibodies derived from these plasmablasts. Blood plasmablasts are elevated in IPAH, remain elevated over time, and produce IgA autoantibodies. Single-cell sequencing of plasmablasts in IPAH revealed repertoires of affinity-matured antibodies with small clonal expansions, consistent with an ongoing autoimmune response. Recombinant antibodies representative of these clonal lineages bound known autoantigen targets and displayed an unexpectedly high degree of polyreactivity. Representative IPAH plasmablast recombinant antibodies stimulated human umbilical vein endothelial cells to produce cytokines and overexpress the adhesion molecule ICAM-1. Together, our results demonstrate an ongoing adaptive autoimmune response involving IgA plasmablasts that produce anti-endothelial cell autoantibodies in IPAH. These antibodies stimulate endothelial cell production of cytokines and adhesion molecules, which may contribute to disease pathogenesis. These findings suggest a role for mucosally-driven autoimmunity and autoimmune injury in the pathogenesis of IPAH. This article is protected by copyright. All rights reserved.

  View details for PubMedID 29369345

• The tyrosine kinase inhibitor imatinib mesylate suppresses uric acid crystal-induced acute gouty arthritis in mice PLOS ONE Reber, L. L., Starkl, P., Balbino, B., Sibilano, R., Gaudenzio, N., Rogalla, S., Sensarn, S., Kang, D., Raghu, H., Sokolove, J., Robinson, W. H., Contag, C. H., Tsai, M., Galli, S. J. 2017; 12 (10): e0185704

  Abstract

  Gouty arthritis is caused by the deposition of monosodium urate (MSU) crystals in joints. Despite many treatment options for gout, there is a substantial need for alternative treatments for patients unresponsive to current therapies. Tyrosine kinase inhibitors have demonstrated therapeutic benefit in experimental models of antibody-dependent arthritis and in rheumatoid arthritis in humans, but to date, the potential effects of such inhibitors on gouty arthritis has not been evaluated. Here we demonstrate that treatment with the tyrosine kinase inhibitor imatinib mesylate (imatinib) can suppress inflammation induced by injection of MSU crystals into subcutaneous air pouches or into the ankle joint of wild type mice. Moreover, imatinib treatment also largely abolished the lower levels of inflammation which developed in IL-1R1-/- or KitW-sh/W-sh mice, indicating that this drug can inhibit IL-1-independent pathways, as well as mast cell-independent pathways, contributing to pathology in this model. Imatinib treatment not only prevented ankle swelling and synovial inflammation when administered before MSU crystals but also diminished these features when administrated after the injection of MSU crystals, a therapeutic protocol more closely mimicking the clinical situation in which treatment occurs after the development of an acute gout flare. Finally, we also assessed the efficiency of local intra-articular injections of imatinib-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles in this model of acute gout. Treatment with low doses of this long-acting imatinib:PLGA formulation was able to reduce ankle swelling in a therapeutic protocol. Altogether, these results raise the possibility that tyrosine kinase inhibitors might have utility in the treatment of acute gout in humans.

  View details for PubMedID 28982129

• Using Flow and Mass Cytometry to Demonstrate Robust Tissue Processing to Query Molecular Heterogeneity in Phase 1 of the Accelerating Medicines Partnership (AMP) - RA Network Wei, K., Rao, D., Zhang, F., Fonseka, C., Slowikowski, K., Keegan, J., Donlin, L. T., Turner, J., McGeachy, M. J., Meednu, N., Lieb, D., Kelly, S., Goodman, S. M., Boyle, D. L., Robinson, W. H., Utz, P. J., Firestein, G. S., Perlman, H., DiCarlo, E. F., Pitzalis, C., Filer, A., Boyce, B., Gravallese, E. M., Nusbaum, C., Lederer, J., Hacohen, N., Gregersen, P., Moreland, L. W., Holers, M., Bykerk, V. P., Raychaudhuri, S., Brenner, M., Anolik, J. H. WILEY. 2017

  View details for Web of Science ID 000411824102322

• Enrichment of malondialdehyde-acetaldehyde antibody in the rheumatoid arthritis joint RHEUMATOLOGY Mikuls, T. R., Duryee, M. J., Rahman, R., Anderson, D. R., Sayles, H. R., Hollins, A., Michaud, K., Wolfe, F., Thiele, G. E., Sokolove, J., Robinson, W. H., Lingampalli, N., Nicholas, A. P., Talmon, G. A., Su, K., Zimmerman, M. C., Klassen, L. W., Thiele, G. M. 2017; 56 (10): 1794–1803

  Abstract

  To characterize the expression of malondialdehdye-acetaldehyde (MAA) adducts and anti-MAA antibody in articular tissues and serum of patients with RA.Paired sera and SF were examined from 29 RA and 13 OA patients. Anti-MAA antibody, RF, ACPA and total immunoglobulin were quantified. SF-serum measures were compared within and between disease groups. The presence and co-localization of MAA, citrulline and select leukocyte antigens in RA and OA synovial tissues were examined using immunohistochemistry.Circulating and SF anti-MAA antibody concentrations were higher in RA vs OA by 1.5- to 5-fold. IgG (P < 0.001), IgM (P = 0.006) and IgA (P = 0.036) anti-MAA antibodies were higher in paired RA SF than serum, differences not observed for total immunoglobulin, RF or ACPA. In RA synovial tissues, co-localization of MAA with citrulline and CD19+ or CD27+ B cells was demonstrated and was much higher in magnitude than MAA or citrulline co-localization with T cells, monocytes, macrophages or dendritic cells (P < 0.01).Anti-MAA antibodies are present in higher concentrations in the RA joint compared with sera, a finding not observed for other disease-related autoantibodies. Co-localization of MAA and citrulline with mature B cells, coupled with the local enrichment of anti-MAA immune responses, implicates MAA-adduct formation in local autoantibody production.

  View details for PubMedID 28957552

• Fibroblast-like Synovial Cell Production of ED-a Fibronectin Contributes to Inflammation in Osteoarthritis Kragstrup, T., Sohn, D., Lepus, C., Onuma, K., Wang, Q., Robinson, W. H., Sokolove, J. WILEY. 2017

  View details for Web of Science ID 000411824104158

• Pharmacodynamic Analysis of Whole Blood Gene Expression over 2 Years in a Phase IIIb Head-to-Head Trial of Abatacept and Adalimumab in Patients with RA Jabado, O., Maldonado, M. A., Schiff, M., Weinblatt, M., Fleischmann, R., Robinson, W. H., Greenfield, A., Connolly, S. E. WILEY. 2017

  View details for Web of Science ID 000411824106372

• Methods for Generating Multiple High-Dimensional Analyses of Cryopreserved Synovial Tissue Developed By the Accelerating Medicines Partnership RA/SLE Network Rao, D., Donlin, L. T., Wei, K., Meednu, N., Turner, J., McGeachy, M. J., Mizoguchi, F., Keegan, J., Lederer, J., Gutierrez-Arcelus, M., Slowikowski, K., Muskat, K., Hillman, J., Rozo, C., Ricker, E., Eisenhaure, T., Lieb, D., Li, S., Browne, E., Nusbaum, C., Robinson, W. H., Kelly, S., Pernis, A. B., Ivashkiv, L., Goodman, S. M., Gravallese, E. M., Holers, M., Hacohen, N., Pitzalis, C., Gregersen, P., Bykerk, V. P., Moreland, L. W., Firestein, G., Raychaudhuri, S., Filer, A., Boyle, D. L., Brenner, M., Anolik, J. H. WILEY. 2017

  View details for Web of Science ID 000411824102323

• STRATEGY FOR ANALYSIS OF DONOR-SPECIFIC ANTIBODY REPERTOIRE: PAIRED IMMUNOGLOBULIN HEAVY AND LIGHT CHAIN ANALYSIS OF INDIVIDUAL MEMORY B CELLS Lucia, M., Gomez, A., Bestard, O., Robinson, W., Martinez, O. WILEY. 2017: 103

  View details for Web of Science ID 000411688501033

• Differential expression of hepatocyte growth factor in patients with systemic sclerosis-associated pulmonary arterial hypertension JOURNAL OF SCLERODERMA AND RELATED DISORDERS Sung, Y. K., Zamanian, R. T., Wagner, C. A., Robinson, W., Steen, V., Chung, L. 2017; 2 (3): 225–30

  View details for DOI 10.5301/jsrd.5000245

  View details for Web of Science ID 000412826600013

• Human Leukocyte Antigen Shared Epitope and Inflammation, Cardiovascular Disease, Cancer, and Mortality Among Postmenopausal Women in the Women's Health Initiative Rheumatoid Arthritis Study AMERICAN JOURNAL OF EPIDEMIOLOGY Talabi, M., Mackey, R. H., Kuller, L. H., Dorman, J. S., Deane, K. D., Robinson, W. H., Walitt, B. T., Chang, Y., Holers, V., Liu, S., Moreland, L. W. 2017; 186 (2): 245–54

  Abstract

  Specific alleles of the human leukocyte antigen (HLA)-DRB1 gene (HLA-DRB1) encode a "shared epitope" (SE) associated with rheumatoid arthritis (RA), especially more severe cyclic-citrullinated peptide antibody-positive (anti-CCP+) RA. We evaluated associations of number of SE alleles (0, 1, or 2) with total and cardiovascular disease (CVD) mortality and incident coronary heart disease (CHD), CVD, and cancer over a mean 8.9 (standard deviation, 3.5) years of follow-up, stratifying by baseline anti-CCP status (positive (+) vs. negative (-)). A longitudinal study, the Women's Health Initiative RA Study (1993-2010), sampled postmenopausal women who reported RA at baseline (1993-1998) or follow-up in the Women's Health Initiative, classified as anti-CCP+ RA (n = 556) or anti-CCP- non-RA (n = 1,070). Among anti-CCP+ RA women, SE alleles were not related to age-adjusted risks of CHD, CVD, or cancer or to total or CVD mortality. Among anti-CCP- non-RA women, age-adjusted hazard ratios for 1 and 2 SE alleles versus 0 SE alleles were 0.41 (95% confidence interval (CI): 0.34, 0.50) and 0.44 (95% CI: 0.27, 0.72), respectively, for CVD; 0.43 (95% CI: 0.37, 0.53) and 0.30 (95% CI: 0.16, 0.64), respectively, for CHD; and 0.62 (95% CI: 0.53, 0.73) and 0.52 (95% CI: 0.33, 0.83), respectively, for cancer. Associations persisted after adjustment for CVD risk factors, joint pain, rheumatoid factor positivity, and inflammatory markers (white blood cell count or cytokine level). In future studies, investigators should evaluate SE associations among anti-CCP- adults without RA and potential mechanisms.

  View details for DOI 10.1093/aje/kwx087

  View details for Web of Science ID 000405869200014

  View details for PubMedID 28459968

  View details for PubMedCentralID PMC5860272

• VP4-and VP7-specific antibodies mediate heterotypic immunity to rotavirus in humans SCIENCE TRANSLATIONAL MEDICINE Nair, N., Feng, N., Blum, L. K., Sanyal, M., Ding, S., Jiang, B., Sen, A., Morton, J. M., He, X., Robinson, W. H., Greenberg, H. B. 2017; 9 (395)

  Abstract

  Human rotaviruses (RVs) are the leading cause of severe diarrhea in young children worldwide. The molecular mechanisms underlying the rapid induction of heterotypic protective immunity to RV, which provides the basis for the efficacy of licensed monovalent RV vaccines, have remained unknown for more than 30 years. We used RV-specific single cell-sorted intestinal B cells from human adults, barcode-based deep sequencing of antibody repertoires, monoclonal antibody expression, and serologic and functional characterization to demonstrate that infection-induced heterotypic immunoglobulins (Igs) primarily directed to VP5*, the stalk region of the RV attachment protein, VP4, are able to mediate heterotypic protective immunity. Heterotypic protective Igs against VP7, the capsid glycoprotein, and VP8*, the cell-binding region of VP4, are also generated after infection; however, our data suggest that homotypic anti-VP7 and non-neutralizing VP8* responses occur more commonly in people. These results indicate that humans can circumvent the extensive serotypic diversity of circulating RV strains by generating frequent heterotypic neutralizing antibody responses to VP7, VP8*, and most often, to VP5* after natural infection. These findings further suggest that recombinant VP5* may represent a useful target for the development of an improved, third-generation, broadly effective RV vaccine and warrants more direct examination.

  View details for PubMedID 28637924

• CCL2/CCR2, but not CCL5/CCR5, mediates monocyte recruitment, inflammation and cartilage destruction in osteoarthritis ANNALS OF THE RHEUMATIC DISEASES Raghu, H., Lepus, C. M., Wang, Q., Wong, H. H., Lingampalli, N., Oliviero, F., Punzi, L., Giori, N. J., Goodman, S. B., Chu, C. R., Sokolove, J. B., Robinson, W. H. 2017; 76 (5)

  Abstract

  While various monocyte chemokine systems are increased in expression in osteoarthritis (OA), the hierarchy of chemokines and chemokine receptors in mediating monocyte/macrophage recruitment to the OA joint remains poorly defined. Here, we investigated the relative contributions of the CCL2/CCR2 versus CCL5/CCR5 chemokine axes in OA pathogenesis.Ccl2-, Ccr2-, Ccl5- and Ccr5-deficient and control mice were subjected to destabilisation of medial meniscus surgery to induce OA. The pharmacological utility of blocking CCL2/CCR2 signalling in mouse OA was investigated using bindarit, a CCL2 synthesis inhibitor, and RS-504393, a CCR2 antagonist. Levels of monocyte chemoattractants in synovial tissues and fluids from patients with joint injuries without OA and those with established OA were investigated using a combination of microarray analyses, multiplexed cytokine assays and immunostains.Mice lacking CCL2 or CCR2, but not CCL5 or CCR5, were protected against OA with a concomitant reduction in local monocyte/macrophage numbers in their joints. In synovial fluids from patients with OA, levels of CCR2 ligands (CCL2, CCL7 and CCL8) but not CCR5 ligands (CCL3, CCL4 and CCL5) were elevated. We found that CCR2+ cells are abundant in human OA synovium and that CCR2+ macrophages line, invade and are associated with the erosion of OA cartilage. Further, blockade of CCL2/CCR2 signalling markedly attenuated macrophage accumulation, synovitis and cartilage damage in mouse OA.Our findings demonstrate that monocytes recruited via CCL2/CCR2, rather than by CCL5/CCR5, propagate inflammation and tissue damage in OA. Selective targeting of the CCL2/CCR2 system represents a promising therapeutic approach for OA.

  View details for DOI 10.1136/annrheumdis-2016-210426

  View details for Web of Science ID 000398387200022

• Nicotine drives neutrophil extracellular traps formation and accelerates collagen-induced arthritis RHEUMATOLOGY Lee, J., Luria, A., Rhodes, C., Raghu, H., Lingampalli, N., Sharpe, O., Rada, B., Sohn, D. H., Robinson, W. H., Sokolove, J. 2017; 56 (4): 644-653

  Abstract

  The aim was to investigate the effects of nicotine on neutrophil extracellular traps (NETs) formation in current and non-smokers and on a murine model of RA.We compared spontaneous and phorbol 12-myristate 13-acetate-induced NETosis between current and non-smokers by DNA release binding. Nicotine-induced NETosis from non-smokers was assessed by DNA release binding, NET-specific (myeloperoxidase (MPO)-DNA complex) ELISA and real-time fluorescence microscopy. We also used immunofluorescent staining to detect nicotinic acetylcholine receptors (nAChRs) on neutrophils and performed a functional analysis to assess the role of nAChRs in nicotine-induced NETosis. Finally, we investigated the effects of systemic nicotine exposure on arthritis severity and NETosis in the CIA mouse model.Neutrophils derived from current smokers displayed elevated levels of spontaneous and phorbol 12-myristate 13-acetate-induced NETosis. Nicotine induced dose-dependent NETosis in ex vivo neutrophils from healthy non-smokers, and co-incubation with ACPA-immune complexes or TNF-α facilitated a synergistic effect on NETosis. Real-time fluorescence microscopy revealed robust formation of NET-like structures in nicotine-exposed neutrophils. Immunofluorescent staining demonstrated the presence of the α7 subunit of the nAChR on neutrophils. Stimulation of neutrophils with an α7-specific nAChR agonist induced NETosis, whereas pretreatment with an nAChR antagonist attenuated nicotine-induced NETosis. Nicotine administration to mice with CIA exacerbated inflammatory arthritis, with higher plasma levels of NET-associated MPO-DNA complex.We demonstrate that nicotine is a potent inducer of NETosis, which may play an important role in accelerating arthritis in the CIA model. This study generates awareness of and the mechanisms by which nicotine-containing products, including e-cigarettes, may have deleterious effects on patients with RA.

  View details for DOI 10.1093/rheumatology/kew449

  View details for Web of Science ID 000398497700023

• Synovial fibroblast-neutrophil interactions promote pathogenic adaptive immunity in rheumatoid arthritis SCIENCE IMMUNOLOGY Carmona-Rivera, C., Carlucci, P. M., Moore, E., Lingampalli, N., Uchtenhagen, H., James, E., Liu, Y., Bicker, K. L., Wahamaa, H., Hoffmann, V., Catrina, A., Thompson, P. R., Buckner, J. H., Robinson, W. H., Fox, D. A., Kaplan, M. J. 2017; 2 (10)

  Abstract

  Rheumatoid arthritis (RA) is characterized by synovial joint inflammation and by development of pathogenic humoral and cellular autoimmunity to citrullinated proteins. Neutrophil extracellular traps (NETs) are a source of citrullinated autoantigens and activate RA synovial fibroblasts (FLS), cells crucial in joint damage. We investigated the molecular mechanisms by which NETs promote proinflammatory phenotypes in FLS, and whether these interactions generate pathogenic anti-citrulline adaptive immune responses. NETs containing citrullinated peptides are internalized by FLS through a RAGE-TLR9 pathway promoting FLS inflammatory phenotype and their upregulation of MHC class II. Once internalized, arthritogenic NET-peptides are loaded into FLS MHC class II and presented to Ag-specific T cells. HLADRB1*0401 transgenic mice immunized with mouse FLS loaded with NETs develop antibodies specific to citrullinated forms of relevant RA autoantigens implicated in RA pathogenesis as well as cartilage damage. These results implicate FLS as mediators in RA pathogenesis, through the internalization and presentation of NET citrullinated peptides to the adaptive immune system leading to pathogenic autoimmunity and cartilage damage.

  View details for PubMedID 28649674

• PKC-epsilon and TLR4 synergistically regulate resistin-mediated inflammation in human macrophages ATHEROSCLEROSIS Zuniga, M. C., Raghuraman, G., Hitchner, E., Weyand, C., Robinson, W., Zhou, W. 2017; 259: 51–59

  Abstract

  Resistin has been associated with atherosclerotic inflammation and cardiovascular complications. We and others have previously shown that PKC-epsilon (PKCε) is involved in resistin-induced smooth muscle cell (VSMC) dysfunction at a high pathological concentration. This study aimed to evaluate the role and potential pathways of resistin at a physiological concentration, in atherosclerosis-related inflammation.Plasma from patients with atherosclerosis was analyzed for resistin concentration. Patients were divided into tertiles based on resistin levels and cytokines were compared between tertiles. Macrophages were then treated with resistin in the presence or absence of PKCε inhibitor and/or TLR4 blocking-antibody, and their inflammatory state was evaluated with ELISA, RT-PCR, immunocytochemistry, and Western blot.We observed significant associations between plasma resistin levels and TNF-α, IL-6, IL-12, MIP-1α, MIP-1β, and CD40L. Our in vitro analyses revealed that resistin activated PKCε via TLR4. This was followed by NF-kB activation and induction of a pro-inflammatory phenotype in macrophages, significantly upregulating CD40, downregulating CD206 and stimulating gene expression and secretion of the inflammatory cytokines, for which we found association in our plasma analysis. Resistin also induced persistent TRAM and CD40L upregulation up to 36 h after resistin treatment. PKCε and TLR4 inhibitors suppressed gene expression to levels similar to control, especially when used in combination.Resistin, at a physiological concentration, exacerbates the inflammatory response of macrophages. PKCε is a key upstream mediator in resistin-induced inflammation that may interact synergistically with TLR4 to promote NF-kB activation, while TRAM is an important signal. PKCε and TRAM may represent novel molecular targets for resistin-associated chronic atherosclerotic inflammation.

  View details for PubMedID 28286252

  View details for PubMedCentralID PMC5395299

• Association of anti-citrullinated protein or peptide antibodies with left ventricular structure and function in rheumatoid arthritis RHEUMATOLOGY Geraldino-Pardilla, L., Russo, C., Sokolove, J., Robinson, W. H., Zartoshti, A., Van Eyk, J., Fert-Bober, J., Lima, J., Giles, J. T., Bathon, J. M. 2017; 56 (4): 534-540

  Abstract

  High levels of ACPAs in RA are associated with more severe arthritis and worse prognosis. However, the role of ACPAs in mediating the increased risk of heart failure in RA remains undefined. We examined whether specific ACPAs were associated with subclinical left ventricular (LV) phenotypes that presage heart failure.Sera from RA patients without clinical cardiovascular disease were assayed for specific ACPAs using a custom Bio-Plex bead assay, and their cross-sectional associations with cardiac magnetic resonance-derived LV measures were evaluated. High ACPA level was defined as ⩾ 75th percentile. Findings were assessed in a second independent RA cohort with an expanded panel of ACPAs and LV measures assessed by 3D-echocardiography.In cohort 1 (n = 76), higher levels of anti-citrullinated fibrinogen 41-60 and anti-citrullinated vimentin antibodies were associated with a 10 and 6% higher adjusted mean LV mass index (LVMI), respectively, compared with lower antibody levels (P < 0.05). In contrast, higher levels of anti-citrullinated biglycan 247-266 were associated with a 13% lower adjusted mean LVMI compared with lower levels (P < 0.001). In cohort 2 (n = 74), the association between ACPAs targeting citrullinated fibrinogen and citrullinated vimentin peptides or protein and LVMI was confirmed: higher anti-citrullinated fibrinogen 556-575 and anti-citrullinated vimentin 58-77 antibody levels were associated with a higher adjusted mean LVMI (19 and 15%, respectively; P < 0.05), but no association with biglycan was found.Higher levels of antibodies targeting citrullinated fibrinogen and vimentin peptides or protein were associated with a higher mean LVMI in both RA cohorts, potentially implicating autoimmune targeting of citrullinated proteins in myocardial remodelling in RA.

  View details for DOI 10.1093/rheumatology/kew436

  View details for Web of Science ID 000398497700007

• Decreased Synovial Inflammation in Atraumatic Hip Microinstability Compared With Femoroacetabular Impingement. Arthroscopy Abrams, G. D., Luria, A., Sampson, J., Madding, R. A., Robinson, W. H., Safran, M. R., Sokolove, J. 2017; 33 (3): 553-558

  Abstract

  To compare the inflammatory profile of hip synovial tissue in those with atraumatic microinstability to patients with femoroacetabular impingement (FAI).Patients with cam and mixed-type FAI (FAI group) and patients with hip instability underwent sampling of the anterolateral synovium. Demographic data, intraoperative measurements, and functional outcome scores (International Hip Outcomes Tool and Short Form-12) were recorded. Cryosections were stained and examined under light microscopy as well as confocal fluorescent microscopy for anti-CD45 (common leukocyte antigen), anti-CD31 (endothelial), and anti-CD68 (macrophage) cell surface markers. A grading system was used to quantify synovitis under light microscopy whereas digital image analysis was used to quantify immunofluorescence staining area. Comparison were made with Student t test, Mann-Whitney U, χ(2), and regression analysis.There were 12 patients in the FAI group and 5 in the instability group. Mean age was not significantly different (P > .05), but there was a significantly greater proportion of females in the instability group versus the FAI group (P < .001). There was a significant correlation (r = 0.653; P = .005) between number of turns needed for 10 mm of distraction and increased synovitis. Synovitis scores also were increased significantly in patients with cam morphology and articular cartilage damage (P = .024) versus those without. Immunohistochemistry did not reveal differences (P > .082) between the instability and FAI groups, but CD68 staining was significantly greater in those with cam morphology and cartilage damage (P < .045). CD45+/CD68- cells were noted in the perivascular area while CD45+/CD68+ cells were noted within the synovial lining in both groups.Increased synovial inflammation was associated with an increased number of turns to achieve joint distraction. Both instability and FAI groups demonstrated baseline levels of synovial inflammation. Synovitis scores also were increased in patients with cartilage damage.An understanding of the molecular and cellular mechanisms behind both hip instability and FAI may lead to novel therapeutic anti-inflammatory therapy, which may serve as an adjunct to treatment of mechanical abnormalities in this conditions.

  View details for DOI 10.1016/j.arthro.2016.09.007

  View details for PubMedID 27939067

• ANTIBODIES TO A SUBSET OF CITRULLINATED PEPTIDE ANTIGENS CORRELATE WITH NEUTROPHIL EXTRACELLULAR TRAP LEVELS IN THE SPUTUM OF SUBJECTS AT-RISK FOR FUTURE RA Demoruelle, M., Purmalek, M., Seto, N. L., Bowers, E., Norris, J. M., Holers, V., Robinson, W. H., Kaplan, M. J., Deane, K. D. BMJ PUBLISHING GROUP. 2017: A93–A94

  View details for DOI 10.1136/annrheumdis-2016-211055.43

  View details for Web of Science ID 000411783100162

• PAD4 Inhibition is Sufficient for the Amelioration of Collagen-Induced Arthritis. Clinical and experimental immunology Willis, V. C., Banda, N. K., Cordova, K. N., Chandra, P. E., Robinson, W. H., Cooper, D. C., Lugo, D., Mehta, G., Taylor, S., Tak, P. P., Prinjha, R. K., Lewis, H. D., Holers, V. M. 2017

  Abstract

  Citrullination of joint proteins by the protein arginine deiminase (PAD) family of enzymes is recognized increasingly as a key process in the pathogenesis of rheumatoid arthritis. This present study was undertaken to explore the efficacy of a novel PAD4-selective inhibitor, GSK199, in the murine collagen-induced arthritis model of rheumatoid arthritis. Mice were dosed daily from the time of collagen immunization with GSK199. Efficacy was assessed against a wide range of end-points, including clinical disease scores, joint histology and immunohistochemistry, serum and joint citrulline levels and quantification of synovial autoantibodies using a proteomic array containing joint peptides. Administration of GSK199 at 30 mg/kg led to significant effects on arthritis, assessed both by global clinical disease activity and by histological analyses of synovial inflammation, pannus formation and damage to cartilage and bone. In addition, significant decreases in complement C3 deposition in both synovium and cartilage were observed robustly with GSK199 at 10 mg/kg. Neither the total levels of citrulline measurable in joint and serum, nor levels of circulating collagen antibodies, were affected significantly by treatment with GSK199 at any dose level. In contrast, a subset of serum antibodies reactive against citrullinated and non-citrullinated joint peptides were reduced with GSK199 treatment. These data extend our previous demonstration of efficacy with the pan-PAD inhibitor Cl-amidine and demonstrate robustly that PAD4 inhibition alone is sufficient to block murine arthritis clinical and histopathological end-points.

  View details for DOI 10.1111/cei.12932

  View details for PubMedID 28128853

  View details for PubMedCentralID PMC5383440

• Identification of KRT16 as a target of an autoantibody response in complex regional pain syndrome EXPERIMENTAL NEUROLOGY Tajerian, M., Hung, V., Khan, H., Lahey, L. J., Sun, Y., Birklein, F., Kraemer, H. H., Robinson, W. H., Kingery, W. S., Clark, J. D. 2017; 287: 14-20

  Abstract

  Using a mouse model of complex regional pain syndrome (CRPS), our goal was to identify autoantigens in the skin of the affected limb.A CRPS-like state was induced using the tibia fracture/cast immobilization model. Three weeks after fracture, hindpaw skin was homogenized, run on 2-d gels, and probed by sera from fracture and control mice. Spots of interest were analyzed by liquid chromatography-mass spectroscopy (LC-MS) and the list of targets validated by examining their abundance and subcellular localization. In order to measure the autoantigenicity of selected protein targets, we quantified the binding of IgM in control and fracture mice sera, as well as in control and CRPS human sera, to the recombinant protein.We show unique binding between fracture skin extracts and fracture sera, suggesting the presence of auto-antigens. LC-MS analysis provided us a list of potential targets, some of which were upregulated after fracture (KRT16, eEF1a1, and PRPH), while others showed subcellular-redistribution and increased membrane localization (ANXA2 and ENO3). No changes in protein citrullination or carbamylation were observed. In addition to increased abundance, KRT16 demonstrated autoantigenicity, since sera from both fracture mice and CRPS patients showed increased autoantibody binding to recombinant kRT16 protein.Pursuing autoimmune contributions to CRPS provides a novel approach to understanding the condition and may allow the development of mechanism-based therapies. The identification of autoantibodies against KRT16 as a biomarker in mice and in humans is a critical step towards these goals, and towards redefining CRPS as having an autoimmune etiology.

  View details for DOI 10.1016/j.expneurol.2016.10.011

  View details for Web of Science ID 000391158000002

• Upregulation of HERV-K is Linked to Immunity and Inflammation in Pulmonary Arterial Hypertension. Circulation Saito, T. n., Miyagawa, K. n., Chen, S. Y., Tamosiuniene, R. n., Wang, L. n., Sharp, O. n., Samayoa, E. n., Harada, D. n., Moonen, J. A., Cao, A. n., Chen, P. I., Hennigs, J. K., Gu, M. n., Li, C. G., Leib, R. D., Li, D. n., Adams, C. M., Del Rosario, P. A., Bill, M. A., Haddad, F. n., Montoya, J. G., Robinson, W. n., Fantl, W. J., Nolan, G. P., Zamanian, R. T., Nicolls, M. R., Chiu, C. Y., Ariza, M. E., Rabinovitch, M. n. 2017

  Abstract

  Background -Immune dysregulation has been linked to occlusive vascular remodeling in pulmonary arterial hypertension (PAH) that is hereditary, idiopathic or associated with other conditions. Circulating autoantibodies, lung perivascular lymphoid tissue and elevated cytokines have been related to PAH pathogenesis but without clear understanding of how these abnormalities are initiated, perpetuated and connected in the progression of disease. We therefore set out to identify specific target antigens in PAH lung immune complexes as a starting point toward resolving these issues to better inform future application of immunomodulatory therapies. Methods -Lung immune complexes were isolated and PAH target antigens were identified by liquid chromatography tandem mass spectrometry (LCMS), confirmed by ELISA, and localized by confocal microscopy. One PAH antigen linked to immunity and inflammation was pursued and a link to PAH pathophysiology was investigated by next generation sequencing, functional studies in cultured monocytes and endothelial cells (EC) and hemodynamic and lung studies in a rat. Results -SAM domain and HD1 domain-containing protein (SAMHD1), an innate immune factor that suppresses HIV replication was identified and confirmed as highly expressed in immune complexes from 16 hereditary and idiopathic PAH vs. 12 control lungs. Elevated SAMHD1 was localized to endothelial cells (EC), perivascular dendritic cells and macrophages and SAMHD1 antibodies were prevalent in tertiary lymphoid tissue. An unbiased screen using metagenomic sequencing related SAMHD1 to increased expression of human endogenous retrovirus K (HERV-K) in PAH vs. control lungs (n=4 each). HERV-K envelope and deoxyuridine triphosphate nucleotidohydrolase (dUTPase) mRNAs were elevated in PAH vs. control lungs (n=10) and proteins were localized to macrophages. HERV-K dUTPase induced SAMHD1 and pro-inflammatory cytokines (e.g., IL6, IL1β and TNFα) in circulating monocytes and pulmonary arterial (PA) EC, and activated B cells. Vulnerability of PAEC to apoptosis was increased by HERV-K dUTPase in an IL6 independent manner. Furthermore, three weekly injections of HERV-K dUTPase induced hemodynamic and vascular changes of pulmonary hypertension in rats (n=8), and elevated IL6. Conclusions -Our study reveals that upregulation of the endogenous retrovirus HERV-K could both initiate and sustain activation of the immune system and cause vascular changes associated with PAH.

  View details for PubMedID 28935667

• Association of Antibodies to Citrullinated Protein Antigens with Blood Pressure in First-Degree Relatives of Rheumatoid Arthritis Patients: The Studies of the Etiology of Rheumatoid Arthritis AMERICAN JOURNAL OF NEPHROLOGY Hughes-Austin, J. M., Gan, R. W., Deane, K. D., Weisman, M. H., Demoruelle, M., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M., Ix, J. H. 2017; 46 (6): 481–87

  Abstract

  Hypertension is more common in patients with rheumatoid arthritis (RA) than in the general population. It is unknown whether hypertension is due to RA-related medications or the disease itself. Therefore, we sought to investigate associations between RA-related autoantibodies, specifically antibodies to citrullinated protein antigens (ACPA) and systolic blood pressure (SBP) and diastolic blood pressure (DBP) in first-degree relatives of RA patients, who were free of RA and RA-related medications. We hypothesized that a greater number of detectable ACPA would be associated with high SBP and DBP, independent of other risk factors in these first-degree relatives.We evaluated associations between ACPA and SBP and DBP in a cross-sectional study of 72 first-degree relatives (defined as parent, child, or sibling) of RA patients. Fifteen ACPA were measured using a Bio-Plex bead-based assay; each was dichotomized as positive/negative based on pre-specified cut-points. Analysis of covariance was used to evaluate associations between ACPA positivity and SBP and DBP, adjusting for age, sex, race, body mass index (BMI), pack-years of smoking, high sensitivity C-reactive protein (hsCRP), and current use of anti-hypertensive medications.Average age was 51 and 69% were women. Mean SBP was 119 ± 18 and DBP was 74 ± 9 mm Hg. Thirty-three (46%) first-degree relatives were positive for ≥1 ACPA; and were younger, had lower BMI, more pack-years of smoking, and higher hsCRP concentrations compared to ACPA negative first-degree relatives. For each additional positive ACPA, SBP was 0.98 ± 0.5 mm Hg (p = 0.05) higher, and DBP was 0.66 ± 0.3 mm Hg (p = 0.04) higher. Anti-cit-fibrinogen A (211-230) positive and anti-cit-filaggrin positive first-degree relatives had 11.5 and 13.9 mm Hg higher SBP (p = 0.02) respectively. Anti-cit-clusterin, cit-filaggrin, and cit-vimentin positive first-degree relatives had 7-8 mm Hg higher DBP (p = 0.03, 0.05, 0.05 respectively), compared to being negative for these individual ACPA. Consistent with associations between ACPA, SBP, and DBP, anti-cyclic citrullinated peptides (anti-CCP2) positive first-degree relatives had 16.4± (p = 0.03) higher SBP and 12.1± mm Hg (p = 0.01) higher DBP than anti-CCP2 negative first-degree relatives.In first-degree relatives without RA, ACPA positivity is associated with higher SBP and DBP. Subclinical autoimmune processes and ACPA may play a role in the vascular changes potentially leading to hypertension prior to RA onset.

  View details for PubMedID 29237149

  View details for PubMedCentralID PMC5788181

• Nicotine drives neutrophil extracellular traps formation and accelerates collagen-induced arthritis. Rheumatology (Oxford, England) Lee, J., Luria, A., Rhodes, C., Raghu, H., Lingampalli, N., Sharpe, O., Rada, B., Sohn, D. H., Robinson, W. H., Sokolove, J. 2016

  Abstract

  The aim was to investigate the effects of nicotine on neutrophil extracellular traps (NETs) formation in current and non-smokers and on a murine model of RA.We compared spontaneous and phorbol 12-myristate 13-acetate-induced NETosis between current and non-smokers by DNA release binding. Nicotine-induced NETosis from non-smokers was assessed by DNA release binding, NET-specific (myeloperoxidase (MPO)-DNA complex) ELISA and real-time fluorescence microscopy. We also used immunofluorescent staining to detect nicotinic acetylcholine receptors (nAChRs) on neutrophils and performed a functional analysis to assess the role of nAChRs in nicotine-induced NETosis. Finally, we investigated the effects of systemic nicotine exposure on arthritis severity and NETosis in the CIA mouse model.Neutrophils derived from current smokers displayed elevated levels of spontaneous and phorbol 12-myristate 13-acetate-induced NETosis. Nicotine induced dose-dependent NETosis in ex vivo neutrophils from healthy non-smokers, and co-incubation with ACPA-immune complexes or TNF-α facilitated a synergistic effect on NETosis. Real-time fluorescence microscopy revealed robust formation of NET-like structures in nicotine-exposed neutrophils. Immunofluorescent staining demonstrated the presence of the α7 subunit of the nAChR on neutrophils. Stimulation of neutrophils with an α7-specific nAChR agonist induced NETosis, whereas pretreatment with an nAChR antagonist attenuated nicotine-induced NETosis. Nicotine administration to mice with CIA exacerbated inflammatory arthritis, with higher plasma levels of NET-associated MPO-DNA complex.We demonstrate that nicotine is a potent inducer of NETosis, which may play an important role in accelerating arthritis in the CIA model. This study generates awareness of and the mechanisms by which nicotine-containing products, including e-cigarettes, may have deleterious effects on patients with RA.

  View details for DOI 10.1093/rheumatology/kew449

  View details for PubMedID 28013195

• Association of anti-citrullinated protein or peptide antibodies with left ventricular structure and function in rheumatoid arthritis. Rheumatology (Oxford, England) Geraldino-Pardilla, L., Russo, C., Sokolove, J., Robinson, W. H., Zartoshti, A., Van Eyk, J., Fert-Bober, J., Lima, J., Giles, J. T., Bathon, J. M. 2016

  Abstract

  High levels of ACPAs in RA are associated with more severe arthritis and worse prognosis. However, the role of ACPAs in mediating the increased risk of heart failure in RA remains undefined. We examined whether specific ACPAs were associated with subclinical left ventricular (LV) phenotypes that presage heart failure.Sera from RA patients without clinical cardiovascular disease were assayed for specific ACPAs using a custom Bio-Plex bead assay, and their cross-sectional associations with cardiac magnetic resonance-derived LV measures were evaluated. High ACPA level was defined as ⩾ 75th percentile. Findings were assessed in a second independent RA cohort with an expanded panel of ACPAs and LV measures assessed by 3D-echocardiography.In cohort 1 (n = 76), higher levels of anti-citrullinated fibrinogen 41-60 and anti-citrullinated vimentin antibodies were associated with a 10 and 6% higher adjusted mean LV mass index (LVMI), respectively, compared with lower antibody levels (P < 0.05). In contrast, higher levels of anti-citrullinated biglycan 247-266 were associated with a 13% lower adjusted mean LVMI compared with lower levels (P < 0.001). In cohort 2 (n = 74), the association between ACPAs targeting citrullinated fibrinogen and citrullinated vimentin peptides or protein and LVMI was confirmed: higher anti-citrullinated fibrinogen 556-575 and anti-citrullinated vimentin 58-77 antibody levels were associated with a higher adjusted mean LVMI (19 and 15%, respectively; P < 0.05), but no association with biglycan was found.Higher levels of antibodies targeting citrullinated fibrinogen and vimentin peptides or protein were associated with a higher mean LVMI in both RA cohorts, potentially implicating autoimmune targeting of citrullinated proteins in myocardial remodelling in RA.

  View details for PubMedID 27994093

• CCL2/CCR2, but not CCL5/CCR5, mediates monocyte recruitment, inflammation and cartilage destruction in osteoarthritis. Annals of the rheumatic diseases Raghu, H., Lepus, C. M., Wang, Q., Wong, H. H., Lingampalli, N., Oliviero, F., Punzi, L., Giori, N. J., Goodman, S. B., Chu, C. R., Sokolove, J. B., Robinson, W. H. 2016

  Abstract

  While various monocyte chemokine systems are increased in expression in osteoarthritis (OA), the hierarchy of chemokines and chemokine receptors in mediating monocyte/macrophage recruitment to the OA joint remains poorly defined. Here, we investigated the relative contributions of the CCL2/CCR2 versus CCL5/CCR5 chemokine axes in OA pathogenesis.Ccl2-, Ccr2-, Ccl5- and Ccr5-deficient and control mice were subjected to destabilisation of medial meniscus surgery to induce OA. The pharmacological utility of blocking CCL2/CCR2 signalling in mouse OA was investigated using bindarit, a CCL2 synthesis inhibitor, and RS-504393, a CCR2 antagonist. Levels of monocyte chemoattractants in synovial tissues and fluids from patients with joint injuries without OA and those with established OA were investigated using a combination of microarray analyses, multiplexed cytokine assays and immunostains.Mice lacking CCL2 or CCR2, but not CCL5 or CCR5, were protected against OA with a concomitant reduction in local monocyte/macrophage numbers in their joints. In synovial fluids from patients with OA, levels of CCR2 ligands (CCL2, CCL7 and CCL8) but not CCR5 ligands (CCL3, CCL4 and CCL5) were elevated. We found that CCR2+ cells are abundant in human OA synovium and that CCR2+ macrophages line, invade and are associated with the erosion of OA cartilage. Further, blockade of CCL2/CCR2 signalling markedly attenuated macrophage accumulation, synovitis and cartilage damage in mouse OA.Our findings demonstrate that monocytes recruited via CCL2/CCR2, rather than by CCL5/CCR5, propagate inflammation and tissue damage in OA. Selective targeting of the CCL2/CCR2 system represents a promising therapeutic approach for OA.

  View details for DOI 10.1136/annrheumdis-2016-210426

  View details for PubMedID 27965260

• Identification of KRT16 as a target of an autoantibody response in complex regional pain syndrome. Experimental neurology Tajerian, M., Hung, V., Khan, H., Lahey, L. J., Sun, Y., Birklein, F., Krämer, H. H., Robinson, W. H., Kingery, W. S., Clark, J. D. 2016; 287: 14-20

  Abstract

  Using a mouse model of complex regional pain syndrome (CRPS), our goal was to identify autoantigens in the skin of the affected limb.A CRPS-like state was induced using the tibia fracture/cast immobilization model. Three weeks after fracture, hindpaw skin was homogenized, run on 2-d gels, and probed by sera from fracture and control mice. Spots of interest were analyzed by liquid chromatography-mass spectroscopy (LC-MS) and the list of targets validated by examining their abundance and subcellular localization. In order to measure the autoantigenicity of selected protein targets, we quantified the binding of IgM in control and fracture mice sera, as well as in control and CRPS human sera, to the recombinant protein.We show unique binding between fracture skin extracts and fracture sera, suggesting the presence of auto-antigens. LC-MS analysis provided us a list of potential targets, some of which were upregulated after fracture (KRT16, eEF1a1, and PRPH), while others showed subcellular-redistribution and increased membrane localization (ANXA2 and ENO3). No changes in protein citrullination or carbamylation were observed. In addition to increased abundance, KRT16 demonstrated autoantigenicity, since sera from both fracture mice and CRPS patients showed increased autoantibody binding to recombinant kRT16 protein.Pursuing autoimmune contributions to CRPS provides a novel approach to understanding the condition and may allow the development of mechanism-based therapies. The identification of autoantibodies against KRT16 as a biomarker in mice and in humans is a critical step towards these goals, and towards redefining CRPS as having an autoimmune etiology.

  View details for DOI 10.1016/j.expneurol.2016.10.011

  View details for PubMedID 27773721

• Associations of Circulating Cytokines and Chemokines With Cancer Mortality in Men With Rheumatoid Arthritis. Arthritis & rheumatology England, B. R., Sokolove, J., Robinson, W. H., Thiele, G. M., Ganti, A. K., Sayles, H., Michaud, K., Caplan, L., Davis, L. A., Cannon, G. W., Sauer, B., Singh, N., Blair Solow, E., Reimold, A. M., Kerr, G. S., Schwab, P., Baker, J. F., Mikuls, T. R. 2016; 68 (10): 2394-2402

  Abstract

  To examine the potential of circulating cytokines and chemokines as biomarkers of cancer mortality risk in patients with rheumatoid arthritis (RA).Male participants in the Veterans Affairs RA registry were followed up from the time of enrollment until death or December 2013. Cytokines and chemokines were measured in banked serum obtained at the time of enrollment, using a bead-based multiplex assay, and a previously developed cytokine score was calculated. Vital status and cause of death were determined through the National Death Index. Associations of cytokines with cancer mortality were examined using multivariable competing-risks regression.Among 1,190 men with RA, 60 cancer deaths (30 of which were attributable to lung cancer) occurred over 5,307 patient-years of follow-up. The patients had a mean age of 64.5 years, had established disease (median duration 8.7 years), were seropositive for rheumatoid factor (81%) or anti-cyclic citrullinated peptide antibody (77%), and frequently had a history of smoking (82% current or former). Seven of 17 analytes examined were individually associated with cancer mortality. The cytokine score was associated with overall cancer (subhazard ratio [SHR] 1.42, 95% confidence interval [95% CI] 1.08-1.85) and lung cancer (SHR 1.86, 95% CI 1.57-2.19) mortality in multivariable analyses. Those in the highest quartile of cytokine scores had a >2-fold increased risk of overall cancer mortality (P = 0.039) and a 6-fold increased risk of lung cancer mortality (P = 0.028) relative to the lowest quartile. A synergistic interaction between current smoking and high cytokine score was observed.Serum cytokines and chemokines are associated with cancer and lung cancer mortality in men with RA, independent of multiple factors including age, smoking status, and prevalent cancer.

  View details for DOI 10.1002/art.39735

  View details for PubMedID 27111000

• Single cell cloning and recombinant monoclonal antibodies generation from RA synovial B cells reveal frequent targeting of citrullinated histones of NETs ANNALS OF THE RHEUMATIC DISEASES Corsiero, E., Bombardieri, M., Carlotti, E., Pratesi, F., Robinson, W., Migliorini, P., Pitzalis, C. 2016; 75 (10): 1866–75

  Abstract

  Rheumatoid arthritis (RA) is characterised by breach of self-tolerance towards citrullinated antigens with generation of anti-citrullinated peptide/proteins antibodies (ACPA). Currently, the nature and source of citrullinated antigens driving the humoral autoimmune response within synovial ectopic lymphoid structures (ELS) is a crucial unknown aspect of RA pathogenesis. Here we characterised the autoreactive B-cell response of lesional B cells isolated from ELS+RA synovium.Single synovial tissue CD19+cells were Fluorescence Activated Cell Sorting (FACS)-sorted and VH/VL Ig genes cloned to generate recombinant monoclonal antibodies (rmAbs) from patients with ELS+/ACPA+RA.RA-rmAbs immunoreactivity analysis provided the following key findings: (1) in a chIP-based array containing 300 autoantigens and in a 'citrullinome' multiplex assay, a strong reactivity against citrullinated histones H2A/H2B (citH2A/H2B) was observed in ∼40% of RA-rmAbs, followed by cit-fibrinogen and cit-vimentin; (2) anti-citH2A/H2B-reactive RA-rmAbs (but not anti-citH2A/H2B negative) selectively recognised neutrophil extracellular traps (NETs) from peripheral blood and/or RA joint neutrophils; (3) anti-citH2A/citH2B and anti-NET immunobinding was dependent on affinity maturation and was completely abrogated following reversion of hypermutated IgVH/VL genes to germline sequences; (4) ELS+ (not ELS-) RA synovial tissues engrafted into Severe Combined ImmunoDeficiency (SCID) mice released human anti-citH2A/citH2B and anti-NET antibodies in association with the intra-graft expression of CXCL13 and lymphotoxin (LT)-β, two master regulators of ELS.We provided novel evidence that B cells differentiated within synovial ELS in the RA joints frequent target deiminated proteins which could be generated during NETosis of RA synovial neutrophils including histones. Thus, NETs could represent a source of citrullinated antigens fuelling the ACPA autoimmune response within the RA synovium.

  View details for PubMedID 26659717

• Sputum Antibodies to Individual Citrullinated Protein/Peptide Antigens Are Elevated in Subjects at-Risk of Future RA and Subjects with Established Disease Bowers, E., Demoruelle, M., Weisman, M., Norris, J. M., Robinson, W. H., Holers, V., Deane, K. D. WILEY. 2016

  View details for Web of Science ID 000417143405143

• Associations of Serum Anti-Citrullinated Proteins and Cytokines with Radiographic Scores in African-American Rheumatoid Arthritis Patients Sun, D., Robinson, W. H., Cui, X., Laufer, V. A., Danila, M. I., Reynolds, R. J., Raman, C., Ledbetter, S., Szalai, A., Bridges, S. WILEY. 2016

  View details for Web of Science ID 000417143400575

• Anti-Periodontal Bacteria Antibody Titers Are Inversely Correlated with ACPA in RA-Free Individuals with Periodontal Disease Compared to Community Controls Weeding, E., Robinson, L., Sokolove, J., Marchesan, J., Offenbacher, S., Robinson, W. H., Demmer, R., Michalowicz, B., Molitor, J. A. WILEY. 2016

  View details for Web of Science ID 000417143400564

• Autoimmunity to Multiple Antigens Is Expanded in at-Risk Family Members Beyond the Disease Specific Patterns of the SLE or RA Proband James, J. A., Bean, K. M., Chen, H., Young, K. A., Bemis, E. A., Seifert, J., Sargent, M., Deane, K. D., Robinson, B., Hafler, D. A., O'Conner, K., Buckner, J. H., Guthridge, J. M., Norris, J. M., Holers, V. WILEY. 2016

  View details for Web of Science ID 000417143401394

• Oral Health and Anti-Citrullinated Peptide Antibodies in Juvenile Idiopathic Arthritis Grevich, S., Lee, P., Ringold, S., Leroux, B., Leahey, H., Yuasa, M., Foster, J., Sokolove, J., Lahey, L., Robinson, W., Newson, J., Stevens, A. WILEY. 2016

  View details for Web of Science ID 000417143403167

• Association of Anti-Citrullinated Peptide Antibodies with Coronary Artery Calcification in Rheumatoid Arthritis. Arthritis care & research Geraldino-Pardilla, L., Giles, J. T., Sokolove, J., Zartoshti, A., Robinson, W. H., Budoff, M., Detrano, R., Bokhari, S., Bathon, J. M. 2016

  Abstract

  Citrullinated proteins have been found within atherosclerotic plaque. However, to date studies evaluating the association between anti-citrullinated peptide antibodies (ACPAs) and imaging measures of atherosclerosis in patients with rheumatoid arthritis (RA) have been limited to citrullinated-fibrinogen or citrullinated-vimentin seroreactivities and rendered contradictory results. Therefore, our objective was to evaluate this association using an extended panel of ACPAs in a larger sample of RA patients without clinical cardiovascular disease (CVD).ACPAs were identified by a custom Bio-Plex bead assay in 270 patients from two independent RA cohorts without clinical CVD consisting of 195 and 75 patients, respectively. Coronary artery calcium (CAC) was assessed by computed tomography as a measure of coronary artery disease.High levels of anti-citrullinated histone 2B antibodies were strongly associated with higher CAC compared with lower antibody levels (p=0.001); this remained significant after adjusting for traditional CV and RA-specific risk factors (p=0.03). No association between levels of ACPAs and CAC progression at 3 years was seen (p=0.09), however the number of progressors was small (n=92).Higher levels of ACPAs targeting cit-histone 2B were associated with higher CAC scores when compared to lower antibody levels, suggesting a potential role for histone citrullination seroreactivity in atherosclerosis. This article is protected by copyright. All rights reserved.

  View details for DOI 10.1002/acr.23106

  View details for PubMedID 27696777

• Associations of Circulating Cytokines and Chemokines With Cancer Mortality in Men With Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY England, B. R., Sokolove, J., Robinson, W. H., Thiele, G. M., Ganti, A. K., Sayles, H., Michaud, K., Caplan, L., Davis, L. A., Cannon, G. W., Sauer, B., Singh, N., Solow, E. B., Reimold, A. M., Kerr, G. S., Schwab, P., Baker, J. F., Mikuls, T. R. 2016; 68 (10): 2394-2402

  Abstract

  To examine the potential of circulating cytokines and chemokines as biomarkers of cancer mortality risk in patients with rheumatoid arthritis (RA).Male participants in the Veterans Affairs RA registry were followed up from the time of enrollment until death or December 2013. Cytokines and chemokines were measured in banked serum obtained at the time of enrollment, using a bead-based multiplex assay, and a previously developed cytokine score was calculated. Vital status and cause of death were determined through the National Death Index. Associations of cytokines with cancer mortality were examined using multivariable competing-risks regression.Among 1,190 men with RA, 60 cancer deaths (30 of which were attributable to lung cancer) occurred over 5,307 patient-years of follow-up. The patients had a mean age of 64.5 years, had established disease (median duration 8.7 years), were seropositive for rheumatoid factor (81%) or anti-cyclic citrullinated peptide antibody (77%), and frequently had a history of smoking (82% current or former). Seven of 17 analytes examined were individually associated with cancer mortality. The cytokine score was associated with overall cancer (subhazard ratio [SHR] 1.42, 95% confidence interval [95% CI] 1.08-1.85) and lung cancer (SHR 1.86, 95% CI 1.57-2.19) mortality in multivariable analyses. Those in the highest quartile of cytokine scores had a >2-fold increased risk of overall cancer mortality (P = 0.039) and a 6-fold increased risk of lung cancer mortality (P = 0.028) relative to the lowest quartile. A synergistic interaction between current smoking and high cytokine score was observed.Serum cytokines and chemokines are associated with cancer and lung cancer mortality in men with RA, independent of multiple factors including age, smoking status, and prevalent cancer.

  View details for DOI 10.1002/art.39735

  View details for Web of Science ID 000384819400007

• Elevated IgA Plasmablast Levels in Subjects at Risk of Developing Rheumatoid Arthritis. Arthritis & rheumatology Kinslow, J. D., Blum, L. K., Deane, K. D., Demoruelle, M. K., Okamoto, Y., Parish, M. C., Kongpachith, S., Lahey, L. J., Norris, J. M., Robinson, W. H., Holers, V. M. 2016; 68 (10): 2372-2383

  Abstract

  The disease process in rheumatoid arthritis (RA) starts years before the clinical diagnosis is made, and elevated levels of disease-specific autoantibodies can be detected during this period. Early responses to known or novel autoantigens likely drive the eventual production of pathogenic autoimmunity. Importantly, the presence of disease-specific autoantibodies can identify individuals who are at high risk of developing RA but who do not currently have arthritis. The goal of the current study was to characterize plasmablasts from individuals at risk of developing RA.We investigated antibody-secreting plasmablasts derived from a well-characterized cohort of individuals who were at risk of developing RA, based on RA-related serum autoantibody positivity, as compared to patients with early (<1 year) seropositive RA as well as healthy control subjects. The plasmablast antibody repertoires of at-risk subjects were analyzed using DNA barcode-based methods with paired heavy- and light-chain gene sequencing. Cells were single-cell sorted, the cell- and plate-specific DNA barcodes were sequentially added, and next-generation sequencing was performed.Total plasmablast levels were similar in the antibody-positive (1%) and control (0.4-1.6%) groups. However, increased frequencies of IgA+ versus IgG+ plasmablasts were observed in the antibody-positive group (39% IgA+ and 37% IgG+) as compared to other groups (1-9% IgA+ and 71-87% IgG+). Paired antibody sequences from antibody-positive subjects revealed cross-isotype clonal families and similar sequence characteristics in the IgA and IgG plasmablast repertoires. Antibody-positive individuals also demonstrated elevated serum levels of IgA isotype anti-cyclic citrullinated peptide 3 antibodies.The IgA plasmablast dominance in these antibody-positive individuals suggests that a subset of RA-related autoantibodies may arise from mucosal immune responses and may be involved in early disease pathogenesis in individuals who are at risk of developing RA.

  View details for DOI 10.1002/art.39771

  View details for PubMedID 27273876

• Synovial Fibroblast-Neutrophil Interactions Promote Pathogenic Adaptive Immunity in Rheumatoid Arthritis Carmona-Rivera, C., Moore, E., Lingampalli, N., Uchtenhagen, H., James, E., Bicker, K. L., Wahamaa, H., Hoffmann, V., Catrina, A. I., Thompson, P., Buckner, J. H., Robinson, W., Fox, D., Kaplan, M. WILEY. 2016

  View details for Web of Science ID 000417143400565

• Increased pretreatment serum IFN-/ ratio predicts non-response to tumour necrosis factor inhibition in rheumatoid arthritis ANNALS OF THE RHEUMATIC DISEASES Muskardin, T. W., Vashisht, P., Dorschner, J. M., Jensen, M. A., Chrabot, B. S., Kern, M., Curtis, J. R., Danila, M. I., Cofield, S. S., Shadick, N., Nigrovic, P. A., St Clair, E. W., Bingham, C. O., Furie, R., Robinson, W., Genovese, M., Striebich, C. C., O'Dell, J. R., Thiele, G. M., Moreland, L. W., Levesque, M., Bridges, S. L., Gregersen, P. K., Niewold, T. B. 2016; 75 (10): 1757-1762

  Abstract

  Studies suggest that circulating type I interferon (IFN) may predict response to biological agents in rheumatoid arthritis (RA). Prediction of response prior to initiating therapy would represent a major advancement.We studied sera from a test set of 32 patients with RA from the Auto-immune Biomarkers Collaborative Network Consortium and a validation set of 92 patients with RA from the Treatment Efficacy and Toxicity in Rheumatoid Arthritis Database and Repository registry. The test set included those with good response or no response to tumour necrosis factor (TNF) inhibitors at 14 weeks by European League Against Rheumatism criteria. The validation set included subjects with good, moderate or no response at 12 weeks. Total serum type I IFN activity, IFN-α and IFN-β activity were measured using a functional reporter cell assay.In the test set, an increased ratio of IFN-β to IFN-α (IFN-β/α activity ratio) in pretreatment serum associated with lack of response to TNF inhibition (p=0.013). Anti-cyclic citrullinated peptide antibody titre and class of TNF inhibitor did not influence this relationship. A receiver-operator curve supported a ratio of 1.3 as the optimal cut-off. In the validation set, subjects with an IFN-β/α activity ratio >1.3 were significantly more likely to have non-response than good response (OR=6.67, p=0.018). The test had 77% specificity and 45% sensitivity for prediction of non-response compared with moderate or good response. Meta-analysis of test and validation sets confirmed strong predictive capacity of IFN-β/α activity ratio (p=0.005).Increased pretreatment serum IFN-β/α ratio strongly associated with non-response to TNF inhibition. This study supports further investigation of serum type I IFN in predicting outcome of TNF inhibition in RA.

  View details for DOI 10.1136/annrheumdis-2015-208001

  View details for PubMedID 26546586

• Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis. Nature reviews. Rheumatology Robinson, W. H., Lepus, C. M., Wang, Q., Raghu, H., Mao, R., Lindstrom, T. M., Sokolove, J. 2016; 12 (10): 580-592

  Abstract

  Osteoarthritis (OA) has long been viewed as a degenerative disease of cartilage, but accumulating evidence indicates that inflammation has a critical role in its pathogenesis. Furthermore, we now appreciate that OA pathogenesis involves not only breakdown of cartilage, but also remodelling of the underlying bone, formation of ectopic bone, hypertrophy of the joint capsule, and inflammation of the synovial lining. That is, OA is a disorder of the joint as a whole, with inflammation driving many pathologic changes. The inflammation in OA is distinct from that in rheumatoid arthritis and other autoimmune diseases: it is chronic, comparatively low-grade, and mediated primarily by the innate immune system. Current treatments for OA only control the symptoms, and none has been FDA-approved for the prevention or slowing of disease progression. However, increasing insight into the inflammatory underpinnings of OA holds promise for the development of new, disease-modifying therapies. Indeed, several anti-inflammatory therapies have shown promise in animal models of OA. Further work is needed to identify effective inhibitors of the low-grade inflammation in OA, and to determine whether therapies that target this inflammation can prevent or slow the development and progression of the disease.

  View details for DOI 10.1038/nrrheum.2016.136

  View details for PubMedID 27539668

• Elevated BMI and antibodies to citrullinated proteins interact to increase rheumatoid arthritis risk and shorten time to diagnosis: A nested case-control study of women in the Nurses' Health Studies. Seminars in arthritis and rheumatism Tedeschi, S. K., Cui, J., Arkema, E. V., Robinson, W. H., Sokolove, J., Lingampalli, N., Sparks, J. A., Karlson, E. W., Costenbader, K. H. 2016

  Abstract

  Overweight/obesity and anti-citrullinated protein antibodies (ACPA) increase rheumatoid arthritis (RA) risk. We investigated the relationship between body mass index (BMI) and ACPA, tested for an interaction between BMI and ACPA for RA risk, and examined effects of BMI and ACPA on time to RA diagnosis.Within the Nurses' Health Studies, blood samples were collected before diagnosis from medical record-confirmed incident RA cases and matched controls. Multiplex assays measured 7 ACPA subtypes (biglycan, clusterin, enolase, fibrinogen, histone 2A, histone 2B, and vimentin). Logistic regression analyses tested the association of BMI and ACPA and for a multiplicative interaction between BMI groups (≥25 vs. <25kg/m(2)) and ACPA positivity (≥2 vs. <2 subtypes), adjusting for age, smoking, alcohol use, and HLA-shared epitope. In case-only analyses, log-rank tests compared time from blood draw to RA onset by cross-classified BMI/ACPA status.Among 255 pre-RA cases and 778 matched controls, 15.7% vs. 2.1% (p<0.001) had ≥2 ACPA and 49.4% vs. 40.2% (p<0.01) were overweight/obese. Continuous BMI was not associated with presence of ≥2 ACPA [OR per kg/m(2) unit BMI: 1.03 (95% CI: 0.97-1.09)]. However, there was a multiplicative interaction between elevated BMI and the presence of ≥2 ACPA for RA risk (p = 0.027). Women with BMI≥25kg/m(2) and ≥2 ACPA had OR 22.7 (95% CI: 6.64-77.72) for RA. Median time to RA differed by BMI/ACPA group (overall log-rank p<0.001) and was shortest among women with ≥2 ACPA and BMI≥25kg/m(2) [45.0 months, IQR: 17.5-72.5] and longest in women with <2 ACPA and BMI<25kg/m(2) [125.0 months, IQR: 72.0-161.0] (pairwise log-rank p = 0.002).Elevated BMI and presence of ACPA interacted to increase RA risk. Time to RA onset was shortest among overweight/obese women with ≥2 ACPA.

  View details for DOI 10.1016/j.semarthrit.2016.09.001

  View details for PubMedID 27939764

• CCL19 as a Chemokine Risk Factor for Posttreatment Lyme Disease Syndrome: a Prospective Clinical Cohort Study. Clinical and vaccine immunology Aucott, J. N., Soloski, M. J., Rebman, A. W., Crowder, L. A., Lahey, L. J., Wagner, C. A., Robinson, W. H., Bechtold, K. T. 2016; 23 (9): 757-766

  Abstract

  Approximately 10% to 20% of patients optimally treated for early Lyme disease develop persistent symptoms of unknown pathophysiology termed posttreatment Lyme disease syndrome (PTLDS). The objective of this study was to investigate associations between PTLDS and immune mediator levels during acute illness and at several time points following treatment. Seventy-six participants with physician-documented erythema migrans and 26 healthy controls with no history of Lyme disease were enrolled. Sixty-four cytokines, chemokines, and inflammatory markers were measured at each visit for a total of 6 visits over 1 year. An operationalized definition of PTLDS incorporating symptoms and functional impact was applied at 6 months and 1 year following treatment completion, and clinical outcome groups were defined as the return-to-health, symptoms-only, and PTLDS groups. Significance analysis of microarrays identified 7 of the 64 immune mediators to be differentially regulated by group. Generalized logit regressions controlling for potential confounders identified posttreatment levels of the T-cell chemokine CCL19 to be independently associated with clinical outcome group. Receiver operating characteristic analysis identified a CCL19 cutoff of >111.67 pg/ml at 1 month following treatment completion to be 82% sensitive and 83% specific for later PTLDS. We speculate that persistently elevated CCL19 levels among participants with PTLDS may reflect ongoing, immune-driven reactions at sites distal to secondary lymphoid tissue. Our findings suggest the relevance of CCL19 both during acute infection and as an immunologic risk factor for PTLDS during the posttreatment phase. Identification of a potential biomarker predictor for PTLDS provides the opportunity to better understand its pathophysiology and to develop early interventions in the context of appropriate and specific clinical information.

  View details for DOI 10.1128/CVI.00071-16

  View details for PubMedID 27358211

• Fractional Third and Fourth Dose of RTS,S/AS01 Malaria Candidate Vaccine: A Phase 2a Controlled Human Malaria Parasite Infection and Immunogenicity Study JOURNAL OF INFECTIOUS DISEASES Regules, J. A., Cicatelli, S. B., Bennett, J. W., Paolino, K. M., Twomey, P. S., Moon, J. E., Kathcart, A. K., Hauns, K. D., Komisar, J. L., Qabar, A. N., Davidson, S. A., Dutta, S., Griffith, M. E., Magee, C. D., Wojnarski, M., Livezey, J. R., Kress, A. T., Waterman, P. E., Jongert, E., Wille-Reece, U., Volkmuth, W., Emerling, D., Robinson, W. H., Lievens, M., Morelle, D., Lee, C. K., Yassin-Rajkumar, B., Weltzin, R., Cohen, J., Paris, R. M., Waters, N. C., Birkett, A. J., Kaslow, D. C., Ballou, W. R., Ockenhouse, C. F., Vekemans, J. 2016; 214 (5): 762-771

  Abstract

  Three full doses of RTS,S/AS01 malaria vaccine provides partial protection against controlled human malaria parasite infection (CHMI) and natural exposure. Immunization regimens, including a delayed fractional third dose, were assessed for potential increased protection against malaria and immunologic responses.In a phase 2a, controlled, open-label, study of healthy malaria-naive adults, 16 subjects vaccinated with a 0-, 1-, and 2-month full-dose regimen (012M) and 30 subjects who received a 0-, 1-, and 7-month regimen, including a fractional third dose (Fx017M), underwent CHMI 3 weeks after the last dose. Plasmablast heavy and light chain immunoglobulin messenger RNA sequencing and antibody avidity were evaluated. Protection against repeat CHMI was evaluated after 8 months.A total of 26 of 30 subjects in the Fx017M group (vaccine efficacy [VE], 86.7% [95% confidence interval [CI], 66.8%-94.6%]; P < .0001) and 10 of 16 in the 012M group (VE, 62.5% [95% CI, 29.4%-80.1%]; P = .0009) were protected against infection, and protection differed between schedules (P = .040, by the log rank test). The fractional dose boosting increased antibody somatic hypermutation and avidity and sustained high protection upon rechallenge.A delayed third fractional vaccine dose improved immunogenicity and protection against infection. Optimization of the RTS,S/AS01 immunization regimen may lead to improved approaches against malaria.NCT01857869.

  View details for DOI 10.1093/infdis/jiw237

  View details for Web of Science ID 000385346400015

  View details for PubMedID 27296848

• Increased inflammation and disease activity among current cigarette smokers with rheumatoid arthritis: a cross-sectional analysis of US veterans. Rheumatology Sokolove, J., Wagner, C. A., Lahey, L. J., Sayles, H., Duryee, M. J., Reimold, A. M., Kerr, G., Robinson, W. H., Cannon, G. W., Thiele, G. M., Mikuls, T. R. 2016: -?

  Abstract

  Cigarette smoking is a major risk factor for RA and has been associated with increased disease severity and lower rates of disease remission. We hypothesized that inflammation and disease activity would be associated with smoking status and this would be related to levels of ACPA.RA patients from the Veterans Affairs RA registry were studied (n = 1466): 76.9% anti-CCP2 positive, 89% male, median age 63 years (interquartile range 57-72), median disease duration 8.45 years (interquartile range 2.8-18). Baseline serum samples were evaluated for levels of anti-CCP2, RF, 19 distinct ACPAs and 17 cytokines. Smoking status at baseline was recorded as current, former or never. The association of smoking status with cytokines, autoantibodies and disease activity (DAS28) was evaluated.Among anti-CCP-positive RA patients, RA-associated cytokines (false-discovery rates q < 0.1%) and DAS28 (P < 0.01) were higher in current smokers compared with former or never smokers. DAS28 and cytokine levels were similar between former and never smokers. In contrast, ACPA concentrations were higher among both current and former smokers compared with never smokers, and levels of ACPA were not associated with DAS28 or cytokine levels.Among anti-CCP2-positive RA patients, current smoking status is associated with elevations in pro-inflammatory cytokines and increased RA disease activity. Similar levels of inflammation and disease activity among former and never smokers suggests that the detrimental effects of smoking could be ameliorated through tobacco cessation. The effect of tobacco cessation on RA disease activity should be evaluated prospectively.

  View details for PubMedID 27477806

• Quality of Life and Performance Status From a Substudy Conducted Within a Prospective Phase 3 Randomized Trial of Concurrent Standard Radiation Versus Accelerated Radiation Plus Cisplatin for Locally Advanced Head and Neck Carcinoma: NRG Oncology RTOG 0129. International journal of radiation oncology, biology, physics Xiao, C., Zhang, Q., Nguyen-Tân, P. F., List, M., Weber, R. S., Ang, K. K., Rosenthal, D., Filion, E. J., Kim, H., Silverman, C., Raben, A., Galloway, T., Fortin, A., Gore, E., Winquist, E., Jones, C. U., Robinson, W., Raben, D., Le, Q., Bruner, D. 2016

  Abstract

  To analyze quality of life (QOL) and performance status (PS) for head and neck cancer (HNC) patients treated on NRG Oncology RTOG 0129 by treatment (secondary outcome) and p16 status, and to examine the association between QOL/PS and survival.Eligible patients were randomized into either an accelerated-fractionation arm or a standard-fractionation arm, and completed the Performance Status Scale for the Head and Neck (PSS-HN), the Head and Neck Radiotherapy Questionnaire (HNRQ), and the Spitzer Quality of Life Index (SQLI) at 8 time points from before treatment to 5 years after treatment.The results from the analysis of area under the curve showed that QOL/PS was not significantly different between the 2 arms from baseline to year after treatment (P ranged from .39 to .98). The results from general linear mixed models further supported the nonsignificant treatment effects until 5 years after treatment (P=.95, .90, and .84 for PSS-HN Diet, Eating, and Speech, respectively). Before treatment and after 1 year after treatment, p16-positive oropharyngeal cancer (OPC) patients had better QOL than did p16-negative patients (P ranged from .0283 to <.0001 for all questionnaires). However, QOL/PS decreased more significantly from pretreatment to the last 2 weeks of treatment in the p16-positive group than in the p16-negative group (P ranged from .0002 to <.0001). Pretreatment QOL/PS was a significant independent predictor of overall survival, progression-free survival, and local-regional failure but not of distant metastasis (P ranged from .0063 to <.0001).The results indicated that patients in both arms may have experienced similar QOL/PS. p16-positive patients had better QOL/PS at baseline and after 1 year of follow-up. Patients presenting with better baseline QOL/PS scores had better survival.

  View details for DOI 10.1016/j.ijrobp.2016.07.020

  View details for PubMedID 27727063

• Proceedings of the 2016 Childhood Arthritis and Rheumatology Research Alliance (CARRA) Scientific Meeting : Toronto, Canada. 14-17 April 2016. Pediatric rheumatology online journal Fotis, L., Shaikh, N., Baszis, K., French, A., Tarr, P., Grevich, S., Lee, P., Ringold, S., Leroux, B., Leahey, H., Yuasa, M., Foster, J., Sokolove, J., Lahey, L., Robinson, W., Newsom, J., Stevens, A., Karasawa, R., Tamaki, M., Tanaka, M., Sato, T., Yudoh, K., Jarvis, J. N., Moncrieffe, H., Bennett, M. F., Tsoras, M., Luyrink, L., Xu, H., Prahalad, S., Morris, P., Dare, J., Nigrovic, P. A., Rosenkranz, M., Becker, M., O’Neil, K. M., Griffin, T., Lovell, D. J., Grom, A. A., Medvedovic, M., Thompson, S. D., Zhu, L., Jiang, K., Wong, L., Buck, M. J., Chen, Y., Moncrieffe, H., Brungs, L., Liu, T., Wang, T., Jarvis, J. N., Alsaeid, K., Alfailakawi, J., Alenezi, H., Alsaeed, H., Beukelman, T., Natter, M., Ilowite, N., Mieszkalski, K., Burrell, G., Best, B., Bristow, H., Carr, S., Dennos, A., Kaufmann, R., Kimura, Y., Schanberg, L., Blier, P. R., Boneparth, A., Wenderfer, S. E., Moorthy, L. N., Radhakrishna, S. M., Sagcal-Gironella, A. C., von Scheven, E., Gedik, K. C., Siddique, S., Aguiar, C. L., Erkan, D., Cohen, E., Lee, Y., Dossett, M., Mehta, D., Davis, R., Gilbert, M., Goilav, B., Meidan, E., Hsu, J., Boneparth, A., Chua, A., Ardoin, S., Wenderfer, S. E., Von Scheven, E., Ruth, N. M., Hui-Yuen, J., Gedik, K. C., Bermudez, L., Cook, A., Imundo, L., Starr, A., Eichenfield, A., Askanase, A., Janow, G., Schanberg, L. E., Setoguchi, S., Hasselblad, V., Mellins, E. D., Schneider, R., Kimura, Y., Kimura, Y., Grevich, S., Beukelman, T., Morgan, E., Graham, T. B., Ibarra, M., Ruas, Y. S., Klein-Gitelman, M., Onel, K., Prahalad, S., Punaro, M., Ringold, S., Toib, D., Van Mater, H., Weiss, J. E., Weiss, P. F., Mieszkalski, K., Schanberg, L. E., Kwok, T. S., Bisaillon, J., Smith, C., Brosseau, L., Stinson, J., Huber, A. M., Duffy, C. M., April, K. T., Lewandowski, L. B., Scott, C., Li, S. C., Torok, K. S., Rabinovich, C. E., Hong, S. D., Becker, M. L., Dedeoglu, F., Ibarra, M. F., Ferguson, P. J., Fuhbrigge, R. C., Stewart, K. G., Pope, E., Laxer, R. M., Mason, T. G., Higgins, G. C., Li, X., Punaro, M. G., Tomlinson, G., Pullenayegum, E., Matelski, J., Schanberg, L., Feldman, B. M., Manthiram, K., Correa, H., Edwards, K., Oberle, E. J., Bayer, M., Co, D. O., Baris, H. E., Chiu, Y., Huber, A., Kim, S., Oberle, E. J., Beukelman, T., Orandi, A. B., Baszis, K. W., Dharnidharka, V., Hoeltzel, M. F., Reed, A., Huber, A., Tomlinson, G., Pullenayegum, E., Matelski, J., Goh, Y. I., Schanberg, L., Feldman, B. M., Schnabel, A., Range, U., Hahn, G., Siepmann, T., Berner, R., Hedrich, C. M., Stevens, B., Torok, K. S., Li, S., Hershey, N., Curran, M., Higgins, G., Moore, K., Rabinovich, E., Stevens, A. M., Stinson, J., Connelly, M., Huber, A., Luca, N., Spiegel, L., Tsimicalis, A., Luca, S., Tajuddin, N., Berard, R., Barsalou, J., Campillo, S., Dancey, P., Duffy, C., Feldman, B., Johnson, N., McGrath, P., Shiff, N., Tse, S., Tucker, L., Victor, C., Stinson, J., Lalloo, C., Harris, L., Cafazzo, J., Spiegel, L., Feldman, B., Luca, N., Laxer, R., Bullock, D. R., Vehe, R. K., Zhang, L., Correll, C. K., Ganguli, S., Shenberger, M., Korumilli, R., Gottlieb, B., Rodriguez, M., de Ranieri, D., Onel, K., Wagner-Weiner, L., Tesher, M., Wojcicki, E. R., Maletta, K. L., Co, D. O., Malloy, M., Thomson, S., Olson, J. C., Wenderfer, S. E., Gilbert, M., Hsu, J., Sule, S., Rubinstein, T. B., Goilav, B., Okamura, D. M., Chua, A., Greenbaum, L. A., Lane, J. C., von Scheven, E., Ardoin, S. P., Ruth, N. M., Woo, J. M., Malloy, M. M., Jegers, J. A., Hahn, D. J., Hintermeyer, M. K., Martinetti, S. M., Heckel, G. R., Roth-Wojcicki, E. L., Co, D. O. 2016; 14 Suppl 1 (Suppl 1): 41

  Abstract

  P1 Serologic evidence of gut-driven systemic inflammation in juvenile idiopathic arthritis Lampros Fotis, Nur Shaikh, Kevin Baszis, Anthony French, Phillip Tarr P2 Oral health and anti-citrullinated peptide antibodies (ACPA) in juvenile idiopathic arthritis Sriharsha Grevich, Peggy Lee, Sarah Ringold, Brian Leroux, Hannah Leahey, Megan Yuasa, Jessica Foster, Jeremy Sokolove, Lauren Lahey, William Robinson, Joshua Newsom, Anne Stevens P3 Novel autoantigens for endothelial cell antibodies in pediatric rheumatic diseases identified by proteomics Rie Karasawa, Mayumi Tamaki, Megumi Tanaka, Toshiko Sato, Kazuo Yudoh, James N. Jarvis P4 Transcriptional profiling reveals monocyte signature associated with JIA patient poor response to methotrexate Halima Moncrieffe, Mark F. Bennett, Monica Tsoras, Lorie Luyrink, Huan Xu, Sampath Prahalad, Paula Morris, Jason Dare, Peter A. Nigrovic, Margalit Rosenkranz, Mara Becker, Kathleen M. O’Neil, Thomas Griffin, Daniel J. Lovell, Alexei A. Grom, Mario Medvedovic, Susan D. Thompson P5 A multi-dimensional genomic map for polyarticular juvenile idiopathic arthritis Lisha Zhu, Kaiyu Jiang, Laiping Wong, Michael J Buck, Yanmin Chen, Halima Moncrieffe, Laura Brungs, Tao Liu, Ting Wang, James N Jarvis P6 Tocilizumab for treatment of children with refractory JIA Khaled Alsaeid, Jasim Alfailakawi, Hamid Alenezi, Hazim Alsaeed P7 Clinical characteristics of the initial patients enrolled in the Childhood Arthritis and Rheumatology Research Alliance (CARRA) Registry Tim Beukelman, Marc Natter, Norm Ilowite, Kelly Mieszkalski, Grendel Burrell, Brian Best, Helen Bristow, Shannon Carr, Anne Dennos, Rachel Kaufmann, Yukiko Kimura, Laura Schanberg P8 Comparative performance of small and large clinical centers in a comprehensive pediatric rheumatology disease registry Peter R Blier P9 Clinical characteristics of children with membranous lupus nephritis: The Childhood Arthritis and Rheumatology Research Alliance Legacy Registry Alexis Boneparth, Scott E. Wenderfer, L. Nandini Moorthy, Suhas M. Radhakrishna, Anna Carmela P. Sagcal-Gironella, Emily von Scheven P10 Rituximab use in pediatric lupus anticoagulant hypoprothrombinemia syndrome - a two center experience Kader Cetin Gedik, Salma Siddique, Cassyanne L. Aguiar, Doruk Erkan P11 Predictors of complementary and alternative medicine use and response in children with musculoskeletal conditions Ezra Cohen, Yvonne Lee, Michelle Dossett, Darshan Mehta, Roger Davis P12 Comparison of pediatric rheumatology and nephrology survey results for the treatment of refractory proliferative lupus nephritis and renal flare in juvenile SLE Mileka Gilbert, Beatrice Goilav, Esra Meidan, Joyce Hsu, Alexis Boneparth, Anabelle Chua, Stacy Ardoin, Scott E. Wenderfer, Emily Von Scheven, Natasha M. Ruth P13 Transitioning lupus patients from pediatric to adult rheumatology Joyce Hui-Yuen, Kader Cetin Gedik, Liza Bermudez, Ashlea Cook, Lisa Imundo, Amy Starr, Andrew Eichenfield, Anca Askanase P14 The systemic juvenile idiopathic arthritis cohort of the Childhood Arthritis & Rheumatology Research Alliance Registry Ginger Janow, Laura E. Schanberg, Soko Setoguchi, Victor Hasselblad, Elizabeth D. Mellins, Rayfel Schneider, Yukiko Kimura, The CARRA Legacy Registry Investigators P15 Results of the pilot study of the Childhood Arthritis and Rheumatology Research Alliance (CARRA) consensus treatment plans for new-onset systemic juvenile idiopathic arthritis Yukiko Kimura, Sriharsha Grevich, Timothy Beukelman, Esi Morgan, T Brent Graham, Maria Ibarra, Yonit Sterba Ruas, Marisa Klein-Gitelman, Karen Onel, Sampath Prahalad, Marilynn Punaro, Sarah Ringold, Dana Toib, Heather Van Mater, Jennifer E. Weiss, Pamela F. Weiss, Kelly Mieszkalski, Laura E. Schanberg P16 A systemic review of pain relief modalities in juvenile idiopathic arthritis: First step in developing a novel decision support intervention Timothy S. H. Kwok, Jacinthe Bisaillon, Christine Smith, Lucie Brosseau, Jennifer Stinson, Adam M. Huber, Ciaran M. Duffy, Karine Toupin April P17 Barriers and facilitators to care retention for pediatric systemic lupus erythematous patients in South Africa: A qualitative study Laura B Lewandowski, Christiaan Scott P18 Evaluating the feasibility of conducting comparative effectiveness studies in juvenile Localized Scleroderma (jLS) Suzanne C. Li, Kathryn S. Torok, C. Egla Rabinovich, Sandy D. Hong, Mara L Becker, Fatma Dedeoglu, Maria F. Ibarra, Polly J Ferguson, Rob C. Fuhbrigge, Katie G. Stewart, Elena Pope, Ronald M. Laxer, Thomas G. Mason, Gloria C. Higgins, Xiaohu Li, Marilynn G. Punaro, George Tomlinson, Eleanor Pullenayegum, John Matelski, Laura Schanberg, Brian M. Feldman P19 Tonsillar histology in patients with periodic fever, aphthous stomatitis, pharyngitis, adenitis (PFAPA) syndrome Kalpana Manthiram, Hernan Correa, Kathryn Edwards P20 Clinical course of juvenile dermatomyositis presenting as skin predominant disease Edward J. Oberle, Michelle Bayer, Dominic O. Co, Hatice Ezgi Baris, Yvonne Chiu, Adam Huber, Susan Kim P21 A Survey of musculoskeletal ultrasound practices of pediatric rheumatologists in North America Edward J Oberle, Timothy Beukelman P22 Assessment, classification and treatment of calcinosis as a complication of juvenile dermatomyositis: A survey of pediatric rheumatologists by the Childhood Arthritis and Rheumatology Research Alliance Amir B. Orandi, Kevin W. Baszis, Vikas Dharnidharka, Mark F. Hoeltzel, for the CARRA JDM Committee P23 CARRA dermatomyositis CTP pilot study Ann Reed, Adam Huber, George Tomlinson, Eleanor Pullenayegum, John Matelski, Y. Ingrid Goh, Laura Schanberg, Brian M. Feldman P24 Unexpectedly high incidences and prolonged disease activity in children with chronic non-bacterial osteomyelitis (CNO) as compared to bacterial osteomyelitis Anja Schnabel, Ursula Range, Gabriele Hahn, Timo Siepmann, Reinhard Berner, Christian Michael Hedrich P25 Juvenile systemic sclerosis cohort within the Childhood Arthritis and Rheumatology Research Alliance (CARRA) Legacy Registry: Follow up characteristics Brandi Stevens, Kathryn S. Torok, Suzanne Li, Nicole Hershey, Megan Curran, Gloria Higgins, Katharine Moore, Egla Rabinovich, Anne M. Stevens, for the CARRA Registry Investigators P26 Development and usability testing of an iPad and desktop psycho-educational game for children with Juvenile Idiopathic Arthritis and their parents Jennifer Stinson, Mark Connelly, Adam Huber, Nadia Luca, Lynn Spiegel, Argerie Tsimicalis, Stephanie Luca, Naweed Tajuddin, Roberta Berard, Julia Barsalou, Sarah Campillo, Paul Dancey, Ciaran Duffy, Brian Feldman, Nicole Johnson, Patrick McGrath, Natalie Shiff, Shirley Tse, Lori Tucker, Charles Victor P27 iCanCopeTM: User-centred design and development of a smartphone app to support self-management for youth with arthritis pain Jennifer Stinson, Chitra Lalloo, Lauren Harris, Joseph Cafazzo, Lynn Spiegel, Brian Feldman, Nadia Luca, Ronald Laxer P28 Accessing pediatric rheumatology care: Despite barriers, few parents prefer telemedicine Danielle R. Bullock, Richard K. Vehe, Lei Zhang, Colleen K. Correll1 P29 Exploration of factors contributing to time to achieve clinically inactive disease (CID) in juvenile idiopathic arthritis (JIA): A preliminary report Suhas Ganguli, Max Shenberger, Ritesh Korumilli, Beth Gottlieb P30 Pediatric rheumatology referral patterns: Presenting complaints of new patients at a large, urban academic center Martha Rodriguez, Deirdre de Ranieri, Karen Onel, Linda Wagner-Weiner, Melissa Tesher P31 Quality improvement (QI) initiatives in childhood systemic lupus erythematosus (cSLE) Elizabeth Roth Wojcicki, Kristyn L. Maletta, Dominic O. Co, Marsha Malloy, Sarah Thomson, Judyann C. Olson P32 Proliferative lupus nephritis in juvenile SLE: Support from the pediatric nephrology community for the definitions of responsiveness and flare in the 2012 consensus treatment plans Scott E. Wenderfer, Mileka Gilbert, Joyce Hsu, Sangeeta Sule, Tamar B. Rubinstein, Beatrice Goilav, Daryl M. Okamura, Annabelle Chua, Laurence A. Greenbaum, Jerome C. Lane, Emily von Scheven, Stacy P. Ardoin, Natasha M. Ruth P33 The steroid taper app: Making of a mobile app Jennifer M. P. Woo, Marsha M. Malloy, James A. Jegers, Dustin J. Hahn, Mary K. Hintermeyer, Stacey M. Martinetti, Gretchen R. Heckel, Elizabeth L. Roth-Wojcicki, Dominic O. Co

  View details for DOI 10.1186/s12969-016-0098-0

  View details for PubMedID 27409414

  View details for PubMedCentralID PMC4943514

• B cell depletion with rituximab in patients with rheumatoid arthritis: Multiplex bead array reveals the kinetics of IgG and IgA antibodies to citrullinated antigens JOURNAL OF AUTOIMMUNITY Cambridge, G., Leandro, M. J., Lahey, L. J., Fairhead, T., Robinson, W. H., Sokolove, J. 2016; 70: 22-30

  Abstract

  The serology of patients with Rheumatoid arthritis (RA) is characterized by persistently raised levels of autoantibodies: Rheumatoid Factors (RhF) against Fc of IgG, and to citrullinated (Cit) protein/peptide sequences: ACPA, recognizing multiple Cit-sequences. B cell depletion therapy based on rituximab delivers good clinical responses in RA patients, particularly in the seropositive group, with responses sometimes lasting beyond the phase of B cell reconstitution. In general, ACPA levels fall following rituximab, but fluctuations with respect to predicting relapse have proved disappointing. In order to identify possible immunodominant specificities within either IgG- or IgA-ACPA we used a Multiplex bead-based array consisting of 30 Cit-peptides/proteins and 22 corresponding native sequences. The kinetics of the serum ACPA response to individual specificities was measured at key points (Baseline, B cell depletion phase, Relapse) within an initial cycle of rituximab therapy in 16 consecutive patients with severe, active RA. All had achieved significant decreases in Disease Activity Scores-28 and maintained B cell depletion in the peripheral blood (<5 CD19+cells/μl) for at least 3 months. At Baseline, mean fluorescence intensity shown by individual IgG- and IgA-ACPA were strongly correlated (R(2) = 0.75; p < 0.0001) but IgA-ACPA were approximately 10-fold lower. Data were Z-normalised in order to compare serial results and antibody classes. At Baseline, a total of 68 IgG- and 51 IgA-ACPA had Z-scores ≥ 1 (above population mean) were identified, with at least one Cit-antigen identified in each serum. ACPA to individual specificities subsequently fluctuated with 3 different patterns. Most 51/68 (75%) IgG- and 48/51 IgA-ACPA (94%) fell between Baseline and Depletion, of which 57% IgG- and 65% IgA-ACPA rebounded pre-Relapse. Interestingly, 17/68 IgG-ACPA (25%) and some IgA-ACPA (3/51; 6%) transiently increased from Baseline, subsequently falling pre-Relapse. Individual responses to particular Cit-epitopes were not linked to particular patterns of fluctuation, but IgG- and IgA-ACPA to individual Cit-antigens often followed similar courses. Some new IgG- and IgA-ACPA, generally to different Cit-antigens however, arose at Relapse in 4 patients. The complexities of the ACPA response after rituximab may therefore reflect its ability to deplete or modify the function of parent B cell clones, which varies between patients. Although relapse following rituximab invariably follows naïve B cell exit from the bone marrow, these studies show that interactions between both 'new' and residual autoreactive memory B cells may be key to resumption of symptoms. The lack of identification of any immunodominant specificity suggests that the process of citrullination, rather than any particular Cit-antigen drives the autoimmune response in RA patients.

  View details for DOI 10.1016/j.jaut.2016.03.010

  View details for PubMedID 27055777

• Association of synovial inflammation and inflammatory mediators with glenohumeral rotator cuff pathology JOURNAL OF SHOULDER AND ELBOW SURGERY Abrams, G. D., Luria, A., Carr, R. A., Rhodes, C., Robinson, W. H., Sokolove, J. 2016; 25 (6): 989-997

  Abstract

  We hypothesized that patients with full-thickness rotator cuff tears would have greater synovial inflammation compared with those without rotator cuff tear pathology, with gene expression relating to histologic findings.Synovial sampling was performed in 19 patients with full-thickness rotator cuff tears (RTC group) and in 11 patients without rotator cuff pathology (control group). Cryosections were stained and examined under light microscopy and confocal fluorescent microscopy for anti-cluster CD45 (common leukocyte antigen), anti-CD31 (endothelial), and anti-CD68 (macrophage) cell surface markers. A grading system was used to quantitate synovitis under light microscopy, and digital image analysis was used to quantify the immunofluorescence staining area. Quantitative polymerase chain reaction was performed for validated inflammatory markers. Data were analyzed with analysis of covariance, Mann-Whitney U, and Spearman rank order testing, with significance set at α = .05.The synovitis score was significantly increased in the RTC group compared with controls. Immunofluorescence demonstrated significantly increased staining for CD31, CD45, and CD68 in the RTC vs control group. CD45+/68- cells were found perivascularly, with CD45+/68+ cells toward the joint lining edge of the synovium. Levels of matrix metalloproteinase-3 (MMP-3) and interleukin-6 were significantly increased in the RTC group, with a positive correlation between the synovitis score and MMP-3 expression.Patients with full-thickness rotator cuff tears have greater levels of synovial inflammation, angiogenesis, and MMP-3 upregulation compared with controls. Gene expression of MMP-3 correlates with the degree of synovitis.

  View details for DOI 10.1016/j.jse.2015.10.011

  View details for PubMedID 26775747

• Expanded IgG lineages from antibody repertoires of melanoma patients to reveal anti-tumor antibodies. Minor, D. R., Defalco, J., Harbell, M., Scholz, A., Manning-Bog, A., Sokolove, J., Robinson, W. H., Emerling, D. AMER SOC CLINICAL ONCOLOGY. 2016

  View details for DOI 10.1200/JCO.2016.34.15_suppl.e23113

  View details for Web of Science ID 000404712500714

• Complement component C1q limits osteoarthritis pathology by regulating macrophage activation Raghu, H., Lingampalli, N., Wang, Q., Wong, H., Lepus, C., Mao, R., Robinson, W. H. AMER ASSOC IMMUNOLOGISTS. 2016

  View details for Web of Science ID 000380288300062

• Use of a MCL-1 inhibitor alone to de-bulk melanoma and in combination to kill melanoma initiating cells. Oncotarget Mukherjee, N., Lu, Y., Almeida, A., Lambert, K., Shiau, C., Su, J., Luo, Y., Fujita, M., Robinson, W. A., Robinson, S. E., Norris, D. A., Shellman, Y. G. 2016

  Abstract

  MCL-1 (BCL-2 family anti-apoptotic protein) is responsible for melanoma's resistance to therapy. Cancer initiating cells also contribute to resistance and relapse from treatments. Here we examined the effects of the MCL-1 inhibitor SC-2001 in killing non melanoma-initiating-cells (bulk of melanoma), and melanoma-initiating-cells (MICs). By itself, SC-2001 significantly kills melanoma cells under monolayer conditions in vitro and in a conventional mouse xenograft model. However, even at high doses (10μM), SC-2001 does not effectively eliminate MICs. In contrast, the combination of SC-2001 with ABT-737 (a BCL-2/BCL-XL/BCL-W inhibitor) significantly decreases ALDH+ cells, disrupts primary spheres, and inhibits the self-renewability of MICs. These results were observed in multiple melanomas, including short term cultures of relapsed tumors from current treatments, independent of the mutation status of BRAF or NRAS. Using a low-cell-number mouse xenograft model, we examined the effects of these treatments on the tumor initiating ability of MIC-enriched cultures. The combination therapy reduces tumor formation significantly compared to either drug alone. Mechanistic studies using shRNA and the CRISPR-Cas9 technology demonstrated that the upregulation of pro-apoptotic proteins NOXA and BIM contribute to the combination-induced cell death. These results indicate that the MCL-1 inhibitor SC-2001 combined with ABT-737 is a promising treatment strategy for targeting melanoma.

  View details for DOI 10.18632/oncotarget.8695

  View details for PubMedID 27086916

• Impact of baseline anti-cyclic citrullinated peptide-2 antibody concentration on efficacy outcomes following treatment with subcutaneous abatacept or adalimumab: 2-year results from the AMPLE trial ANNALS OF THE RHEUMATIC DISEASES Sokolove, J., Schiff, M., Fleischmann, R., Weinblatt, M. E., Connolly, S. E., Johnsen, A., Zhu, J., Maldonado, M. A., Patel, S., Robinson, W. H. 2016; 75 (4): 709-714

  Abstract

  To examine whether baseline anti-cyclic citrullinated peptide-2 (CCP2) antibody status and concentration correlated with clinical outcomes in patients treated with abatacept or adalimumab on background methotrexate (MTX) in the 2-year AMPLE (Abatacept versus adaliMumab comParison in bioLogic-naïvE rheumatoid arthritis subjects with background MTX) study.In this exploratory analysis, anti-CCP2 antibody concentration was measured at baseline, and antibody-positive patients were divided into equal quartiles, Q1-Q4, representing increasing antibody concentrations. Clinical outcomes analysed by baseline anti-CCP2 status and quartile included change from baseline in disease activity and disability and remission rates.Baseline characteristics were generally comparable across quartiles and treatment groups. In both treatment groups, anti-CCP2 antibody-negative patients responded less well than antibody-positive patients. At year 2, improvements in disease activity and disability and remission rates were similar across Q1-Q3, but were numerically higher in Q4 in the abatacept group; in contrast, treatment effects were similar across all quartiles in the adalimumab group.In AMPLE, baseline anti-CCP2 positivity was associated with a better response for abatacept and adalimumab. Patients with the highest baseline anti-CCP2 antibody concentrations had better clinical response with abatacept than patients with lower concentrations, an association that was not observed with adalimumab.NCT00929864.

  View details for DOI 10.1136/annrheumdis-2015-207942

  View details for PubMedID 26359449

• Anti-Insulin Immune Responses Are Detectable in Dogs with Spontaneous Diabetes PLOS ONE Kim, J., Furrow, E., Ritt, M. G., Utz, P. J., Robinson, W. H., Yu, L., Eckert, A., Stuebner, K., O'Brien, T. D., Steinman, L., Modiano, J. F. 2016; 11 (3)

  Abstract

  Diabetes mellitus occurs spontaneously in dogs. Although canine diabetes shares many features with human type-1 diabetes, there are differences that have cast doubt on the immunologic origin of the canine disease. In this study, we examined whether peripheral immune responses directed against islet antigens were present in dogs with diabetes. Routine diagnostics were used to confirm diabetic status, and serum samples from dogs with (N = 15) and without (N = 15) diabetes were analyzed for the presence of antibodies against islet antigens (insulin, glutamic acid decarboxylase, insulinoma-associated protein tyrosine phosphatase, and islet beta-cell zinc cation efflux transporter) using standard radioassays. Interferon-γ production from peripheral blood T cells stimulated by porcine insulin and by human insulin was tested using Elispot assays. Anti-insulin antibodies were detectable in a subset of diabetic dogs receiving insulin therapy. Pre-activated T cells and incipient insulin-reactive T cells in response to porcine or human insulin were identified in non-diabetic dogs and in dogs with diabetes. The data show that humoral and cellular anti-insulin immune responses are detectable in dogs with diabetes. This in turn provides support for the potential to ethically use dogs with diabetes to study the therapeutic potential of antigen-specific tolerance.

  View details for DOI 10.1371/journal.pone.0152397

  View details for PubMedID 27031512

• The interleukin-20 receptor axis in early rheumatoid arthritis: novel links between disease-associated autoantibodies and radiographic progression ARTHRITIS RESEARCH & THERAPY Kragstrup, T. W., Greisen, S. R., Nielsen, M. A., Rhodes, C., Stengaard-Pedersen, K., Hetland, M. L., Horslev-Petersen, K., Junker, P., Ostergaard, M., Hvid, M., Vorup-Jensen, T., Robinson, W. H., Sokolove, J., Deleuran, B. 2016; 18

  Abstract

  Rheumatoid arthritis (RA) is often characterized by the presence of rheumatoid factor, anti-citrullinated protein antibodies, and bone erosions. Current therapies can compromise immunity, leading to risk of infection. The interleukin-20 receptor (IL-20R) axis comprising IL-19, IL-20, and IL-24 and their shared receptors activates tissue homeostasis processes but not the immune system. Consequently, modulation of the IL-20R axis may not lead to immunosuppression, making it an interesting drug target. We evaluated the role of the IL-20R axis in RA and associations between plasma cytokine levels and clinical disease.Plasma IL-19, IL-20, and IL-24 levels were measured in early RA patients during a treat-to-target strategy by enzyme-linked immunosorbent assays. The IL-20R1 and IL-22R1 levels in paired peripheral blood mononuclear cells and synovial fluid mononuclear cells from a different cohort of RA patients were evaluated by flow cytometry and confocal microscopy. Monocytes/macrophages were stimulated with heat-aggregated human immunoglobulin immune complexes and immune complexes containing citrullinated fibrinogen, and osteoclasts were incubated with IL-19, IL-20, and IL-24.The plasma concentrations of IL-20 and IL-24 (but not IL-19) were increased in early RA patients compared with healthy controls (both P < 0.002) and decreased after 6 months of treatment (both P < 0.0001). The expression of IL-22R1 (but not IL-20R1) was increased on monocytes from RA synovial fluid compared with monocytes from both RA and healthy control peripheral blood. The plasma concentrations of IL-20 and IL-24 were increased in rheumatoid factor and anti-citrullinated protein antibody positive compared with negative early RA patients (all P < 0.0001). Immune complexes stimulated the production of the IL-20R cytokines by monocytes/macrophages. Increased baseline plasma concentrations of IL-20 and IL-24 were associated with Sharp-van der Heijde score progression after 24 months (Spearman's rho = 0.19 and 0.26, both P < 0.05) in the early RA patients. The IL-22R1 was expressed by osteoclast precursors and in multinucleated osteoclasts. IL-20 and IL-24 increased the secretion of monocyte chemoattractant protein 1 by these cells.This study suggests that IL-20 and IL-24 link RA-associated autoantibodies with radiographic progression via the IL-22R1. Modulation of this axis holds promise as feasible anti-erosive treatment modalities in seropositive RA.

  View details for DOI 10.1186/s13075-016-0964-7

  View details for Web of Science ID 000372114700001

  View details for PubMedCentralID PMC4788924

• Biomarkers to guide clinical therapeutics in rheumatology? CURRENT OPINION IN RHEUMATOLOGY Robinson, W. H., Mao, R. 2016; 28 (2): 168-175

  Abstract

  The use of biomarkers in rheumatology can help identify disease risk, improve diagnosis and prognosis, target therapy, assess response to treatment, and further our understanding of the underlying pathogenesis of disease. Here, we discuss the recent advances in biomarkers for rheumatic disorders, existing impediments to progress in this field, and the potential of biomarkers to enable precision medicine and thereby transform rheumatology.Although significant challenges remain, progress continues to be made in biomarker discovery and development for rheumatic diseases. The use of next-generation technologies, including large-scale sequencing, proteomic technologies, metabolomic technologies, mass cytometry, and other single-cell analysis and multianalyte analysis technologies, has yielded a slew of new candidate biomarkers. Nevertheless, these biomarkers still require rigorous validation and have yet to make their way into clinical practice and therapeutic development. This review focuses on advances in the biomarker field in the last 12 months as well as the challenges that remain.Better biomarkers, ideally mechanistic ones, are needed to guide clinical decision making in rheumatology. Although the use of next-generation techniques for biomarker discovery is making headway, it is imperative that the roadblocks in our search for new biomarkers are overcome to enable identification of biomarkers with greater diagnostic and predictive utility. Identification of biomarkers with robust diagnostic and predictive utility would enable precision medicine in rheumatology.

  View details for DOI 10.1097/BOR.0000000000000250

  View details for Web of Science ID 000369424300012

  View details for PubMedID 26720904

  View details for PubMedCentralID PMC4770786

• Multiplexed autoantigen microarrays identify HLA as a key driver of anti-desmoglein and -non-desmoglein reactivities in pemphigus PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Sajda, T., Hazelton, J., Patel, M., Seiffert-Sinha, K., Steinman, L., Robinson, W., Haab, B. B., Sinha, A. A. 2016; 113 (7): 1859-1864

  Abstract

  Patients with pemphigus vulgaris (PV) harbor antibodies reactive against self-antigens expressed at the surface of keratinocytes, primarily desmoglein (Dsg) 3 and, to a lesser extent, Dsg1. Conventionally, only antibodies targeting these molecules have been thought to contribute to disease pathogenesis. This notion has been challenged by a growing pool of evidence that suggests that antibodies toward additional targets may play a role in disease. The aims of this study were to (i) establish high-throughput protein microarray technology as a method to investigate traditional and putative autoantibodies (autoAbs) in PV and (ii) use multiplexed protein array technology to define the scope and specificity of the autoAb response in PV. Our analysis demonstrated significant IgG reactivity in patients with PV toward the muscarinic acetylcholine receptor subtypes 3, 4, and 5 as well as thyroid peroxidase. Furthermore, we found that healthy first- and second-degree relatives of patients with PV express autoAbs toward desmoglein and non-Dsg targets. Our analysis also identified genetic elements, particularly HLA, as key drivers of autoAb expression. Finally, we show that patients with PV exhibit significantly reduced IgM reactivity toward disease-associated antigens relative to controls. The use of protein microarrays to profile the autoAb response in PV advanced the current understanding of disease and provided insight into the complex relationship between genetics and disease development.

  View details for DOI 10.1073/pnas.1525448113

  View details for PubMedID 26831096

• Identification of Candidate Tolerogenic CD8(+) T Cell Epitopes for Therapy of Type 1 Diabetes in the NOD Mouse Model. Journal of diabetes research Yu, C., Burns, J. C., Robinson, W. H., Utz, P. J., Ho, P. P., Steinman, L., Frey, A. B. 2016; 2016: 9083103-?

  Abstract

  Type 1 diabetes is an autoimmune disease in which insulin-producing pancreatic islet β cells are the target of self-reactive B and T cells. T cells reactive with epitopes derived from insulin and/or IGRP are critical for the initiation and maintenance of disease, but T cells reactive with other islet antigens likely have an essential role in disease progression. We sought to identify candidate CD8(+) T cell epitopes that are pathogenic in type 1 diabetes. Proteins that elicit autoantibodies in human type 1 diabetes were analyzed by predictive algorithms for candidate epitopes. Using several different tolerizing regimes using synthetic peptides, two new predicted tolerogenic CD8(+) T cell epitopes were identified in the murine homolog of the major human islet autoantigen zinc transporter ZnT8 (aa 158-166 and 282-290) and one in a non-β cell protein, dopamine β-hydroxylase (aa 233-241). Tolerizing vaccination of NOD mice with a cDNA plasmid expressing full-length proinsulin prevented diabetes, whereas plasmids encoding ZnT8 and DβH did not. However, tolerizing vaccination of NOD mice with the proinsulin plasmid in combination with plasmids expressing ZnT8 and DβH decreased insulitis and enhanced prevention of disease compared to vaccination with the plasmid encoding proinsulin alone.

  View details for DOI 10.1155/2016/9083103

  View details for PubMedID 27069933

• Development of a Multiantigen Panel for Improved Detection of Borrelia burgdorferi Infection in Early Lyme Disease JOURNAL OF CLINICAL MICROBIOLOGY Lahey, L. J., Panas, M. W., Mao, R., Delanoy, M., Flanagan, J. J., Binder, S. R., Rebman, A. W., Montoya, J. G., Soloski, M. J., Steere, A. C., Dattwyler, R. J., Arnaboldi, P. M., Aucott, J. N., Robinson, W. H. 2015; 53 (12): 3834-3841

  Abstract

  The current standard for laboratory diagnosis of Lyme disease in the United States is serologic detection of antibodies against Borrelia burgdorferi. The Centers for Disease Control and Prevention recommends a two-tiered testing algorithm; however, this scheme has limited sensitivity for detecting early Lyme disease. Thus, there is a need to improve diagnostics for Lyme disease at the early stage, when antibiotic treatment is highly efficacious. We examined novel and established antigen markers to develop a multiplex panel that identifies early infection using the combined sensitivity of multiple markers while simultaneously maintaining high specificity by requiring positive results for two markers to designate a positive test. Ten markers were selected from our initial analysis of 62 B. burgdorferi surface proteins and synthetic peptides by assessing binding of IgG and IgM to each in a training set of Lyme disease patient samples and controls. In a validation set, this 10-antigen panel identified a higher proportion of early-Lyme-disease patients as positive at the baseline or posttreatment visit than two-tiered testing (87.5% and 67.5%, respectively; P < 0.05). Equivalent specificities of 100% were observed in 26 healthy controls. Upon further analysis, positivity on the novel 10-antigen panel was associated with longer illness duration and multiple erythema migrans. The improved sensitivity and comparable specificity of our 10-antigen panel compared to two-tiered testing in detecting early B. burgdorferi infection indicates that multiplex analysis, featuring the next generation of markers, could advance diagnostic technology to better aid clinicians in diagnosing and treating early Lyme disease.

  View details for DOI 10.1128/JCM.02111-15

  View details for Web of Science ID 000367532800020

  View details for PubMedCentralID PMC4652118

• Development of a Multiantigen Panel for Improved Detection of Borrelia burgdorferi Infection in Early Lyme Disease. Journal of clinical microbiology Lahey, L. J., Panas, M. W., Mao, R., Delanoy, M., Flanagan, J. J., Binder, S. R., Rebman, A. W., Montoya, J. G., Soloski, M. J., Steere, A. C., Dattwyler, R. J., Arnaboldi, P. M., Aucott, J. N., Robinson, W. H. 2015; 53 (12): 3834-41

  Abstract

  The current standard for laboratory diagnosis of Lyme disease in the United States is serologic detection of antibodies against Borrelia burgdorferi. The Centers for Disease Control and Prevention recommends a two-tiered testing algorithm; however, this scheme has limited sensitivity for detecting early Lyme disease. Thus, there is a need to improve diagnostics for Lyme disease at the early stage, when antibiotic treatment is highly efficacious. We examined novel and established antigen markers to develop a multiplex panel that identifies early infection using the combined sensitivity of multiple markers while simultaneously maintaining high specificity by requiring positive results for two markers to designate a positive test. Ten markers were selected from our initial analysis of 62 B. burgdorferi surface proteins and synthetic peptides by assessing binding of IgG and IgM to each in a training set of Lyme disease patient samples and controls. In a validation set, this 10-antigen panel identified a higher proportion of early-Lyme-disease patients as positive at the baseline or posttreatment visit than two-tiered testing (87.5% and 67.5%, respectively; P < 0.05). Equivalent specificities of 100% were observed in 26 healthy controls. Upon further analysis, positivity on the novel 10-antigen panel was associated with longer illness duration and multiple erythema migrans. The improved sensitivity and comparable specificity of our 10-antigen panel compared to two-tiered testing in detecting early B. burgdorferi infection indicates that multiplex analysis, featuring the next generation of markers, could advance diagnostic technology to better aid clinicians in diagnosing and treating early Lyme disease.

  View details for DOI 10.1128/JCM.02111-15

  View details for PubMedID 26447113

  View details for PubMedCentralID PMC4652118

• Absence of keratin 8 or 18 promotes antimitochondrial autoantibody formation in aging male mice FASEB JOURNAL Toivola, D. M., Habtezion, A., Misiorek, J. O., Zhang, L., Nystrom, J. H., Sharpe, O., Robinson, W. H., Kwan, R., Omary, M. B. 2015; 29 (12): 5081-5089

  Abstract

  Human mutations in keratin 8 (K8) and keratin 18 (K18), the intermediate filament proteins of hepatocytes, predispose to several liver diseases. K8-null mice develop chronic liver injury and fragile hepatocytes, dysfunctional mitochondria, and Th2-type colitis. We tested the hypothesis that autoantibody formation accompanies the liver damage that associates with K8/K18 absence. Sera from wild-type control, K8-null, and K18-null mice were analyzed by immunoblotting and immunofluorescence staining of cell and mouse tissue homogenates. Autoantibodies to several antigens were identified in 81% of K8-null male mice 8 mo or older. Similar autoantibodies were detected in aging K18-null male mice that had a related liver phenotype but normal colon compared with K8-null mice, suggesting that the autoantibodies are linked to liver rather than colonic disease. However, these autoantibodies were not observed in nontransgenic mice subjected to 4 chronic injury models. The autoantigens are ubiquitous and partition with mitochondria. Mass spectrometry and purified protein analysis identified, mitochondrial HMG-CoA synthase, aldehyde dehydrogenase, and catalase as the primary autoantigens, and glutamate dehydrogenase and epoxide hydrolase-2 as additional autoantigens. Therefore, absence of the hepatocyte keratins results in production of anti-mitochondrial autoantibodies (AMA) that recognize proteins involved in energy metabolism and oxidative stress, raising the possibility that AMA may be found in patients with keratin mutations that associate with liver and other diseases.

  View details for DOI 10.1096/fj.14-269795

  View details for PubMedID 26399787

• Local Joint Inflammation and Histone Citrullination in a Murine Model of the Transition From Preclinical Autoimmunity to Inflammatory Arthritis ARTHRITIS & RHEUMATOLOGY Sohn, D. H., Rhodes, C., Onuma, K., Zhao, X., Sharpe, O., Gazitt, T., Shiao, R., Fert-Bober, J., Cheng, D., Lahey, L. J., Wong, H. H., Van Eyk, J., Robinson, W. H., Sokolove, J. 2015; 67 (11): 2877-2887

  Abstract

  Anti-citrullinated protein antibodies (ACPAs) are characteristic of rheumatoid arthritis (RA). However, their presence years before the onset of clinical RA is perplexing. Although multiple putative citrullinated antigens have been identified, no studies have demonstrated the specific capacity of these antigens to initiate inflammatory arthritis. This study was undertaken to recapitulate the transition from preclinical to clinical RA and to demonstrate the capacity of local citrullination to facilitate this transition.We performed proteomic analysis of activated human neutrophils to identify citrullinated proteins, including those targeted as part of the RA immune response. Using enzyme-linked immunosorbent assay, we compared RA and osteoarthritis synovial fluid for levels of citrullinated histone H2B and its immune complex. Using macrophage activation assays, we assessed the effect of histone citrullination on immunostimulatory capacity and evaluated the stimulatory capacity of native and citrullinated H2B immune complexes. Finally, we assessed the potential for anti-citrullinated H2B antibodies to mediate arthritis in vivo.We identified robust targeting of neutrophil-derived citrullinated histones by the ACPA immune response. More than 90% of the RA patients had anti-citrullinated H2B antibodies. Histone citrullination increased innate immunostimulatory capacity, and immune complexes containing citrullinated histones activated macrophage cytokine production and propagated neutrophil activation. Finally, we demonstrated that immunization with H2B was arthritogenic, but only in the setting of underlying articular inflammation.Our findings indicate that citrullinated histones, specifically citrullinated H2B, are an antigenic target of the ACPA immune response. Furthermore, local generation of citrullinated antigen during low-grade articular inflammation provides a mechanistic model for the conversion from preclinical autoimmunity to inflammatory arthritis.

  View details for DOI 10.1002/art.39283

  View details for PubMedID 26227989

• DECADE IN REVIEW-TECHNOLOGY Technological advances transforming rheumatology NATURE REVIEWS RHEUMATOLOGY Robinson, W. H., Mao, R. 2015; 11 (11): 626–U14

  View details for DOI 10.1038/nrrheum.2015.137

  View details for Web of Science ID 000364041400003

  View details for PubMedID 26439404

  View details for PubMedCentralID PMC4769868

• Circulating Cytokine/Chemokine Concentrations Predict Cancer Mortality in Men with Rheumatoid Arthritis England, B. R., Sokolove, J., Robinson, W. H., Thiele, G. M., Ganti, A. K., Sayles, H., Michaud, K., Caplan, L., Davis, L. A., Cannon, G. W., Sauer, B., Solow, E., Reimold, A., Kerr, G. S., Schwab, P., Baker, J. F., Mikuls, T. R. WILEY-BLACKWELL. 2015

  View details for Web of Science ID 000370860204784

• The IL-20 Receptor Axis in Early Rheumatoid Arthritis: Novel Inflammation-Independent Links Between Autoantibody Positivity and Radiographic Progression Kragstrup, T., Greisen, S., Nielsen, M., Rhodes, C., Stengaard-Pedersen, K., Hetland, M., Horslev-Petersen, K., Junker, P., Ostergaard, M., Hvid, M., Vorup-Jensen, T., Robinson, W. H., Sokolove, J., Deleuran, B. WILEY-BLACKWELL. 2015

  View details for Web of Science ID 000370860201538

• Associations of Serum Anti-Malondialdehyde-Acetaldehyde (MAA) Antibodies with Cardiovascular and Respiratory Mortality in Men with Rheumatoid Arthritis England, B. R., Thiele, G. M., Duryee, M. J., Klassen, L., Sokolove, J., Robinson, W. H., Anderson, D., Sayles, H., Michaud, K., Caplan, L., Davis, L. A., Cannon, G. W., Sauer, B., Solow, E., Reimold, A., Kerr, G. S., Schwab, P., Baker, J. F., Mikuls, T. R. WILEY-BLACKWELL. 2015

  View details for Web of Science ID 000370860202834

• B Cell Depletion with Rituximab in Patients with Rheumatoid Arthritis: Multiplex Bead Array Reveals Kinetics of IgG and IgA Autoantibodies to Citrullinated Antigens Cambridge, G., Lahey, L. J., Leandro, M. J., Robinson, W. H., Sokolove, J. WILEY-BLACKWELL. 2015

  View details for Web of Science ID 000370860203048

• Increased IgA Plasmablasts Are Associated with Rheumatoid Arthritis-Related Autoantibodies in the Absence of Inflammatory Arthritis Kinslow, J., Blum, L., Deane, K. D., Demoruelle, K., Parish, M. C., Kongpachith, S., Lahey, L. J., Norris, J. M., Robinson, W. H., Holers, V. WILEY-BLACKWELL. 2015

  View details for Web of Science ID 000370860202358

• A Distinct Class of Antibodies May Be an Indicator of Gray Matter Autoimmunity in Early and Established Relapsing Remitting Multiple Sclerosis Patients ASN NEURO Ligocki, A. J., Rivas, J. R., Rounds, W. H., Guzman, A. A., Li, M., Spadaro, M., Lahey, L., Chen, D., Henson, P. M., Graves, D., Greenberg, B. M., Frohman, E. M., Ward, E. S., Robinson, W., Meinl, E., White, C. L., Stowe, A. M., Monson, N. L. 2015; 7 (5)

  Abstract

  *These authors contributed equally to the work in this manuscript.We have previously identified a distinct class of antibodies expressed by B cells in the cerebrospinal fluid (CSF) of early and established relapsing remitting multiple sclerosis (RRMS) patients that is not observed in healthy donors. These antibodies contain a unique pattern of mutations in six codons along VH4 antibody genes that we termed the antibody gene signature (AGS). In fact, patients who have such B cells in their CSF are identified as either having RRMS or developing RRMS in the future. As mutations in antibody genes increase antibody affinity for particular antigens, the goal for this study was to investigate whether AGS(+) antibodies bind to brain tissue antigens. Single B cells were isolated from the CSF of 10 patients with early or established RRMS. We chose 32 of these B cells that expressed antibodies enriched for the AGS for further study. We generated monoclonal full-length recombinant human antibodies (rhAbs) and used both immunological assays and immunohistochemistry to investigate the capacity of these AGS(+) rhAbs to bind brain tissue antigens. AGS(+) rhAbs did not recognize myelin tracts in the corpus callosum. Instead, AGS(+) rhAbs recognized neuronal nuclei and/or astrocytes, which are prevalent in the cortical gray matter. This pattern was unique to the AGS(+) antibodies from early and established RRMS patients, as AGS(+) antibodies from an early neuromyelitis optica patient did not display the same reactivity. Prevalence of CSF-derived B cells expressing AGS(+) antibodies that bind to these cell types may be an indicator of gray matter-directed autoimmunity in early and established RRMS patients.

  View details for DOI 10.1177/1759091415609613

  View details for PubMedID 26489686

• Rheumatoid Arthritis, Anti-Cyclic Citrullinated Peptide Positivity, and Cardiovascular Disease Risk in the Women's Health Initiative. Arthritis & rheumatology (Hoboken, N.J.) Mackey, R. H., Kuller, L. H., Deane, K. D., Walitt, B. T., Chang, Y. F., Holers, V. M., Robinson, W. H., Tracy, R. P., Hlatky, M. A., Eaton, C. B., Liu, S., Freiberg, M. S., Talabi, M. B., Schelbert, E. B., Moreland, L. W. 2015; 67 (9): 2311-22

  Abstract

  To evaluate the incidence of cardiovascular disease (CVD) morbidity and mortality over the course of 10 years among the more than 160,000 postmenopausal women in the Women's Health Initiative (WHI) in relation to self-reported rheumatoid arthritis (RA), taking disease-modifying antirheumatic drugs (DMARDs), anti-cyclic citrullinated peptide (anti-CCP) positivity, rheumatoid factor (RF) positivity, CVD risk factors, joint pain, and inflammation (white blood cell count and interleukin-6 levels).Anti-CCP and RF were measured in a sample of WHI participants with self-reported RA (n = 9,988). RA was classified as self-reported RA plus anti-CCP positivity and/or taking DMARDs. Anti-CCP-negative women with self-reported RA and not taking DMARDs were classified as having "unverified RA."Age-adjusted rates of coronary heart disease (CHD), stroke, CVD, fatal CVD, and total mortality were higher in women with RA than in women with no reported RA, with multivariable-adjusted hazard ratios of 1.46 (95% confidence interval [95% CI] 1.17-1.83) for CHD and 2.55 (95% CI 1.86-3.51) for fatal CVD. Among women with RA, anti-CCP positivity and RF positivity were not significantly associated with higher risk of any outcomes, despite slightly higher risk of death for those who were anti-CCP positive than for those who were anti-CCP negative. Joint pain severity and CVD risk factors were strongly associated with CVD risk, even in women with no reported RA. CVD incidence was increased in women with RA versus women with no reported RA at almost all risk factor levels, except for low levels of joint pain or inflammation. Among women with RA, inflammation was more strongly associated with fatal CVD and total mortality than with CHD or CVD.Among postmenopausal women, RA was associated with 1.5-2.5-fold higher CVD risk. CVD risk was strongly associated with CVD risk factors, joint pain severity, and inflammation, but not with anti-CCP positivity or RF positivity.

  View details for DOI 10.1002/art.39198

  View details for PubMedID 25988241

  View details for PubMedCentralID PMC4551571

• Rheumatoid Arthritis, Anti-Cyclic Citrullinated Peptide Positivity, and Cardiovascular Disease Risk in the Women's Health Initiative ARTHRITIS & RHEUMATOLOGY Mackey, R. H., Kuller, L. H., Deane, K. D., Walitt, B. T., Chang, Y., Holers, V. M., Robinson, W. H., Tracy, R. P., Hlatky, M. A., Eaton, C. B., Liu, S., Freiberg, M. S., Talabi, M. B., Schelbert, E. B., Moreland, L. W. 2015; 67 (9): 2311-2322

  Abstract

  To evaluate the incidence of cardiovascular disease (CVD) morbidity and mortality over the course of 10 years among the more than 160,000 postmenopausal women in the Women's Health Initiative (WHI) in relation to self-reported rheumatoid arthritis (RA), taking disease-modifying antirheumatic drugs (DMARDs), anti-cyclic citrullinated peptide (anti-CCP) positivity, rheumatoid factor (RF) positivity, CVD risk factors, joint pain, and inflammation (white blood cell count and interleukin-6 levels).Anti-CCP and RF were measured in a sample of WHI participants with self-reported RA (n = 9,988). RA was classified as self-reported RA plus anti-CCP positivity and/or taking DMARDs. Anti-CCP-negative women with self-reported RA and not taking DMARDs were classified as having "unverified RA."Age-adjusted rates of coronary heart disease (CHD), stroke, CVD, fatal CVD, and total mortality were higher in women with RA than in women with no reported RA, with multivariable-adjusted hazard ratios of 1.46 (95% confidence interval [95% CI] 1.17-1.83) for CHD and 2.55 (95% CI 1.86-3.51) for fatal CVD. Among women with RA, anti-CCP positivity and RF positivity were not significantly associated with higher risk of any outcomes, despite slightly higher risk of death for those who were anti-CCP positive than for those who were anti-CCP negative. Joint pain severity and CVD risk factors were strongly associated with CVD risk, even in women with no reported RA. CVD incidence was increased in women with RA versus women with no reported RA at almost all risk factor levels, except for low levels of joint pain or inflammation. Among women with RA, inflammation was more strongly associated with fatal CVD and total mortality than with CHD or CVD.Among postmenopausal women, RA was associated with 1.5-2.5-fold higher CVD risk. CVD risk was strongly associated with CVD risk factors, joint pain severity, and inflammation, but not with anti-CCP positivity or RF positivity.

  View details for DOI 10.1002/art.39198

  View details for Web of Science ID 000360379500004

  View details for PubMedCentralID PMC4551571

• Vascular calcifications on hand radiographs in rheumatoid arthritis and associations with autoantibodies, cardiovascular risk factors and mortality RHEUMATOLOGY Solow, E. B., Yu, F., Thiele, G. M., Sokolove, J., Robinson, W. H., Pruhs, Z. M., Michaud, K. D., Erickson, A. R., Sayles, H., Kerr, G. S., Gaffo, A. L., Caplan, L., Davis, L. A., Cannon, G. W., Reimold, A. M., Baker, J., Schwab, P., Anderson, D. R., Mikuls, T. R. 2015; 54 (9): 1587-1595

  Abstract

  To examine whether vascular calcifications on hand films in RA might aid in determining mortality risk.Hand radiographs from 906 RA patients were scored as positive or negative for vascular calcifications. Patient characteristics associated with vascular calcifications were assessed using multivariable logistic regression, and associations with mortality were examined using Cox proportional hazards regression. Cytokines and multiplex ACPA were measured in both groups.A total of 99 patients (11%) demonstrated radiographic vascular calcifications. Factors independently associated with vascular calcifications included diabetes [odds ratio (OR) 2.85; 95% CI 1.43, 5.66], cardiovascular disease at enrolment (OR 2.48; 95% CI 1.01, 6.09), prednisone use (OR 1.90; 95% CI 1.25, 2.91), current smoking (OR 0.06; 95% CI 0.01, 0.23) and former smoking (OR 0.36; 95% CI 0.27, 0.48) vs never smoking. In cytokine and ACPA subtype analysis, IL-4 and anti-citrullinated apolipoprotein E were significantly increased in patients with vascular calcifications in fully adjusted multivariable models. After multivariable adjustment, vascular calcifications were associated with an increase in all-cause mortality (hazard ratio 1.41; 95% CI 1.12, 1.78; P = 0.004).Vascular calcifications on hand radiographs were independently associated with increased all-cause mortality in RA. Mechanisms underpinning the associations of IL-4 and select ACPA with vascular calcifications and their utility as biomarkers predictive of cardiovascular disease risk in RA merit further study.

  View details for DOI 10.1093/rheumatology/kev027

  View details for Web of Science ID 000362848500009

  View details for PubMedID 25854268

  View details for PubMedCentralID PMC4536858

• Mass cytometry analysis shows that a novel memory phenotype B cell is expanded in multiple myeloma. Cancer immunology research Hansmann, L., Blum, L., Ju, C., Liedtke, M., Robinson, W. H., Davis, M. M. 2015; 3 (6): 650-660

  Abstract

  It would be very beneficial if the status of cancers could be determined from a blood specimen. However, peripheral blood leukocytes are very heterogeneous between individuals, and thus high-resolution technologies are likely required. We used cytometry by time-of-flight and next-generation sequencing to ask whether a plasma cell cancer (multiple myeloma) and related precancerous states had any consistent effect on the peripheral blood mononuclear cell phenotypes of patients. Analysis of peripheral blood samples from 13 cancer patients, 9 precancer patients, and 9 healthy individuals revealed significant differences in the frequencies of the T-cell, B-cell, and natural killer-cell compartments. Most strikingly, we identified a novel B-cell population that normally accounts for 4.0% ± 0.7% (mean ± SD) of total B cells and is up to 13-fold expanded in multiple myeloma patients with active disease. This population expressed markers previously associated with both memory (CD27(+)) and naïve (CD24(lo)CD38(+)) phenotypes. Single-cell immunoglobulin gene sequencing showed polyclonality, indicating that these cells are not precursors to the myeloma, and somatic mutations, a characteristic of memory cells. SYK, ERK, and p38 phosphorylation responses, and the fact that most of these cells expressed isotypes other than IgM or IgD, confirmed the memory character of this population, defining it as a novel type of memory B cells.

  View details for DOI 10.1158/2326-6066.CIR-14-0236-T

  View details for PubMedID 25711758

  View details for PubMedCentralID PMC4457663

• Mass Cytometry Analysis Shows That a Novel Memory Phenotype B Cell Is Expanded in Multiple Myeloma CANCER IMMUNOLOGY RESEARCH Hansmann, L., Blum, L., Ju, C., Liedtke, M., Robinson, W. H., Davis, M. M. 2015; 3 (6): 650-660

  Abstract

  It would be very beneficial if the status of cancers could be determined from a blood specimen. However, peripheral blood leukocytes are very heterogeneous between individuals, and thus high-resolution technologies are likely required. We used cytometry by time-of-flight and next-generation sequencing to ask whether a plasma cell cancer (multiple myeloma) and related precancerous states had any consistent effect on the peripheral blood mononuclear cell phenotypes of patients. Analysis of peripheral blood samples from 13 cancer patients, 9 precancer patients, and 9 healthy individuals revealed significant differences in the frequencies of the T-cell, B-cell, and natural killer-cell compartments. Most strikingly, we identified a novel B-cell population that normally accounts for 4.0% ± 0.7% (mean ± SD) of total B cells and is up to 13-fold expanded in multiple myeloma patients with active disease. This population expressed markers previously associated with both memory (CD27(+)) and naïve (CD24(lo)CD38(+)) phenotypes. Single-cell immunoglobulin gene sequencing showed polyclonality, indicating that these cells are not precursors to the myeloma, and somatic mutations, a characteristic of memory cells. SYK, ERK, and p38 phosphorylation responses, and the fact that most of these cells expressed isotypes other than IgM or IgD, confirmed the memory character of this population, defining it as a novel type of memory B cells.

  View details for DOI 10.1158/2326-6066.CIR-14-0236-T

  View details for Web of Science ID 000357430000010

  View details for PubMedID 25711758

  View details for PubMedCentralID PMC4457663

• CD11c-mediated deletion of Flip promotes autoreactivity and inflammatory arthritis NATURE COMMUNICATIONS Huang, Q., Perlman, H., Birkett, R., Doyle, R., Fang, D., Haines, G. K., Robinson, W., Datta, S., Huang, Z., Li, Q., Phee, H., Pope, R. M. 2015; 6

  Abstract

  Dendritic cells (DCs) are critical for immune homeostasis. To target DCs, we generated a mouse line with Flip deficiency in cells that express cre under the CD11c promoter (CD11c-Flip-KO). CD11c-Flip-KO mice spontaneously develop erosive, inflammatory arthritis, resembling rheumatoid arthritis, which is dramatically reduced when these mice are crossed with Rag(-/-) mice. The CD8α(+) DC subset is significantly reduced, along with alterations in NK cells and macrophages. Autoreactive CD4(+) T cells and autoantibodies specific for joint tissue are present, and arthritis severity correlates with the number of autoreactive CD4(+) T cells and plasmablasts in the joint-draining lymph nodes. Reduced T regulatory cells (Tregs) inversely correlate with arthritis severity, and the transfer of Tregs ameliorates arthritis. This KO line identifies a model that will permit in depth interrogation of the pathogenesis of rheumatoid arthritis, including the role of CD8α(+) DCs and other cells of the immune system.

  View details for DOI 10.1038/ncomms8086

  View details for PubMedID 25963626

• Large-scale sequencing of antibody responses to Borrelia burgdorferi infection Blum, L., Lahey, L., Kongpachith, S., Ju, C., Aucott, J., Soloski, M., Robinson, W. AMER ASSOC IMMUNOLOGISTS. 2015

  View details for Web of Science ID 000379404502124

• Malondialdehyde-Acetaldehyde Adducts and Anti-Malondialdehyde-Acetaldehyde Antibodies in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Thiele, G. M., Duryee, M. J., Anderson, D. R., Klassen, L. W., Mohring, S. M., Young, K. A., Benissan-Messan, D., Sayles, H., Dusad, A., Hunter, C. D., Sokolove, J., Robinson, W. H., O'Dell, J. R., Nicholas, A. P., Tuma, D. J., Mikuls, T. R. 2015; 67 (3): 645-655

  Abstract

  Malondialdehyde-acetaldehyde (MAA) adducts are a product of oxidative stress associated with tolerance loss in several disease states. This study was undertaken to investigate the presence of MAA adducts and circulating anti-MAA antibodies in patients with rheumatoid arthritis (RA).Synovial tissue from patients with RA and patients with osteoarthritis (OA) were examined for the presence of MAA-modified and citrullinated proteins. Anti-MAA antibody isotypes were measured in RA patients (n = 1,720) and healthy controls (n = 80) by enzyme-linked immunosorbent assay. Antigen-specific anti-citrullinated protein antibodies (ACPAs) were measured in RA patients using a multiplex antigen array. Anti-MAA isotype concentrations were compared in a subset of RA patients (n = 80) and matched healthy controls (n = 80). Associations of anti-MAA antibody isotypes with disease characteristics, including ACPA positivity, were examined in all RA patients.Expression of MAA adducts was increased in RA synovial tissue compared to OA synovial tissue, and colocalization with citrullinated proteins was found. Increased levels of anti-MAA antibody isotypes were observed in RA patients compared to controls (P < 0.001). Among RA patients, anti-MAA antibody isotypes were associated with seropositivity for ACPAs and rheumatoid factor (P < 0.001) in addition to select measures of disease activity. Higher anti-MAA antibody concentrations were associated with a greater number of positive antigen-specific ACPA analytes (expressed at high titer) (P < 0.001) and a higher ACPA score (P < 0.001), independent of other covariates.MAA adduct formation is increased in RA and appears to result in robust antibody responses that are strongly associated with ACPAs. These results support speculation that MAA formation may be a cofactor that drives tolerance loss, resulting in the autoimmune responses characteristic of RA.

  View details for DOI 10.1002/art.38969

  View details for Web of Science ID 000350300400009

  View details for PubMedID 25417811

• Sequencing the functional antibody repertoire-diagnostic and therapeutic discovery NATURE REVIEWS RHEUMATOLOGY Robinson, W. H. 2015; 11 (3): 171-182

  Abstract

  The development of high-throughput DNA sequencing technologies has enabled large-scale characterization of functional antibody repertoires, a new method of understanding protective and pathogenic immune responses. Important parameters to consider when sequencing antibody repertoires include the methodology, the B-cell population and clinical characteristics of the individuals analysed, and the bioinformatic analysis. Although focused sequencing of immunoglobulin heavy chains or complement determining regions can be utilized to monitor particular immune responses and B-cell malignancies, high-fidelity analysis of the full-length paired heavy and light chains expressed by individual B cells is critical for characterizing functional antibody repertoires. Bioinformatic identification of clonal antibody families and recombinant expression of representative members produces recombinant antibodies that can be used to identify the antigen targets of functional immune responses and to investigate the mechanisms of their protective or pathogenic functions. Integrated analysis of coexpressed functional genes provides the potential to further pinpoint the most important antibodies and clonal families generated during an immune response. Sequencing antibody repertoires is transforming our understanding of immune responses to autoimmunity, vaccination, infection and cancer. We anticipate that antibody repertoire sequencing will provide next-generation biomarkers, diagnostic tools and therapeutic antibodies for a spectrum of diseases, including rheumatic diseases.

  View details for DOI 10.1038/nrrheum.2014.220

  View details for Web of Science ID 000350667900007

  View details for PubMedID 25536486

  View details for PubMedCentralID PMC4382308

• Identification of anticitrullinated protein antibody reactivities in a subset of anti-CCP-negative rheumatoid arthritis: association with cigarette smoking and HLA-DRB1 'shared epitope' alleles. Annals of the rheumatic diseases Wagner, C. A., Sokolove, J., Lahey, L. J., Bengtsson, C., Saevarsdottir, S., Alfredsson, L., Delanoy, M., Lindstrom, T. M., Walker, R. P., Bromberg, R., Chandra, P. E., Binder, S. R., Klareskog, L., Robinson, W. H. 2015; 74 (3): 579-586

  Abstract

  A hallmark of rheumatoid arthritis (RA) is the development of autoantibodies targeting proteins that contain citrulline. Anticitrullinated protein antibodies (ACPAs) are currently detected by the commercial cyclic citrullinated peptide (CCP) assay, which uses a mix of cyclised citrullinated peptides as an artificial mimic of the true antigen(s). To increase the sensitivity of ACPA detection and dissect ACPA specificities, we developed a multiplex assay that profiles ACPAs by measuring their reactivity to the citrullinated peptides and proteins derived from RA joint tissue.We created a bead-based, citrullinated antigen array to profile ACPAs. This custom array contains 16 citrullinated peptides and proteins detected in RA synovial tissues. We used the array to profile ACPAs in sera from a cohort of patients with RA and other non-inflammatory arthritides, as well as sera from an independent cohort of RA patients for whom data were available on carriage of HLA-DRB1 'shared epitope' (SE) alleles and history of cigarette smoking.Our multiplex assay showed that at least 10% of RA patients who tested negative in the commercial CCP assay possessed ACPAs. Carriage of HLA-DRB1 SE alleles and a history of cigarette smoking were associated with an increase in ACPA reactivity-in anti-CCP(+) RA and in a subset of anti-CCP(-) RA.Our multiplex assay can identify ACPA-positive RA patients missed by the commercial CCP assay, thus enabling greater diagnostic sensitivity. Further, our findings suggest that cigarette smoking and possession of HLA-DRB1 SE alleles contribute to the development of ACPAs in anti-CCP(-) RA.

  View details for DOI 10.1136/annrheumdis-2013-203915

  View details for PubMedID 24297382

• Associations of toll-like receptor (TLR)-4 single nucleotide polymorphisms and rheumatoid arthritis disease progression: An observational cohort study INTERNATIONAL IMMUNOPHARMACOLOGY Davis, M. L., LeVan, T. D., Yu, F., Sayles, H., Sokolove, J., Robinson, W., Michaud, K., Thiele, G. M., Mikuls, T. R. 2015; 24 (2): 346-352

  Abstract

  To examine the associations of toll-like receptor (TLR)-4 single nucleotide polymorphisms (SNPs) with disease progression in rheumatoid arthritis (RA).A total of 1188 RA patients were genotyped for TLR4 SNPs (rs1927911, rs11536878, and rs4986790). Measures of disease activity were examined, including Disease Activity Score-28 (DAS28), Multidimensional Health Assessment Questionnaire (MD-HAQ), Clinical Disease Activity Index (CDAI), and Simplified Disease Activity Index (SDAI). Genetic associations with these longitudinal measures were examined using generalized estimating equations in both univariate and multivariate analyses. Analyses were then stratified by antigen specific anti-citrullinated peptide antibody (ACPA) status including antibody to citrullinated fibrinogen and citrullinated histone H2B.Disease activity measures progressed less over time in the homozygous minor allele group of rs1927911 including DAS28 (p<0.001), CDAI (p=0.008), and MD-HAQ (p=0.015) in univariate analysis and DAS28, CDAI and SDAI in multivariate analysis. Disease activity progression among those homozygous for the minor allele tended to be lower in the groups with positive ACPA though major differences by autoantibody status were not identified. There were no associations of TLR4 rs11536878 and rs4986790 SNPs with RA disease activity progression.In this population, TLR4 rs1927911 genotypes are associated with disease activity independent of other covariates.

  View details for DOI 10.1016/j.intimp.2014.12.030

  View details for PubMedID 25573402

• Alveolar Bone Loss Is Associated With Circulating Anti-Citrullinated Protein Antibody (ACPA) in Patients With Rheumatoid Arthritis JOURNAL OF PERIODONTOLOGY Gonzalez, S. M., Payne, J. B., Yu, F., Thiele, G. M., Erickson, A. R., Johnson, P. G., Schmid, M. J., Cannon, G. W., Kerr, G. S., Reimold, A. M., Sokolove, J., Robinson, W. H., Mikuls, T. R. 2015; 86 (2): 222-231

  Abstract

  This study examines: 1) alveolar bone loss (ABL), a hallmark of periodontitis, in anti-citrullinated protein antibody (ACPA)-positive rheumatoid arthritis (RA) patients versus control patients with osteoarthritis (OA); and 2) the association of ABL with RA disease activity and ACPA concentrations, including multiple antigen-specific ACPA.This multicenter case-control study includes 617 patients diagnosed with RA (n = 287) or OA (n = 330). Panoramic radiographs were taken; patients were categorized into low, moderate, or high tertiles based on mean percentage ABL. Serum ACPA was measured using second-generation anticyclic citrullinated peptide enzyme-linked immunosorbent assay and a multiplex platform to assess distinct antigen-specific ACPA. A generalized linear mixed model for binary data was used to compare stratified ABL in RA versus OA patients. Associations of moderate and high ABL (versus low) with RA disease activity and severity measures were examined using multivariate regression. Antigen-specific ACPA responses were compared among ABL tertiles using significance analysis of microarrays.ACPA-positive patients with RA had a significantly higher mean percentage of sites with ABL >20% compared with patients with OA (P = 0.03). After multivariate adjustment, greater ABL was significantly associated with higher serum ACPA concentration (P = 0.004), 28-joint Disease Activity Score (P = 0.023), health assessment questionnaire disability (P = 0.05), tender joint count (P = 0.02) and joint space narrowing scores (P = 0.05) among patients with RA. ACPAs targeting citrullinated vimentin and histone were significantly higher in moderate and high ABL groups versus low, regardless of smoking status (q <0.1%).Greater ABL was associated with higher ACPA, consistent with findings at articular sites. ACPA targeting could provide novel insight into important linkages between RA and periodontitis.

  View details for DOI 10.1902/jop.2014.140425

  View details for Web of Science ID 000349380500008

  View details for PubMedID 25299390

• The interleukin-20 receptor axis in early rheumatoid arthritis: novel links between disease-associated autoantibodies and radiographic progression. Arthritis research & therapy Kragstrup, T. W., Greisen, S. R., Nielsen, M. A., Rhodes, C., Stengaard-Pedersen, K., Hetland, M. L., Hørslev-Petersen, K., Junker, P., Østergaard, M., Hvid, M., Vorup-Jensen, T., Robinson, W. H., Sokolove, J., Deleuran, B. 2015; 18: 61-?

  Abstract

  Rheumatoid arthritis (RA) is often characterized by the presence of rheumatoid factor, anti-citrullinated protein antibodies, and bone erosions. Current therapies can compromise immunity, leading to risk of infection. The interleukin-20 receptor (IL-20R) axis comprising IL-19, IL-20, and IL-24 and their shared receptors activates tissue homeostasis processes but not the immune system. Consequently, modulation of the IL-20R axis may not lead to immunosuppression, making it an interesting drug target. We evaluated the role of the IL-20R axis in RA and associations between plasma cytokine levels and clinical disease.Plasma IL-19, IL-20, and IL-24 levels were measured in early RA patients during a treat-to-target strategy by enzyme-linked immunosorbent assays. The IL-20R1 and IL-22R1 levels in paired peripheral blood mononuclear cells and synovial fluid mononuclear cells from a different cohort of RA patients were evaluated by flow cytometry and confocal microscopy. Monocytes/macrophages were stimulated with heat-aggregated human immunoglobulin immune complexes and immune complexes containing citrullinated fibrinogen, and osteoclasts were incubated with IL-19, IL-20, and IL-24.The plasma concentrations of IL-20 and IL-24 (but not IL-19) were increased in early RA patients compared with healthy controls (both P < 0.002) and decreased after 6 months of treatment (both P < 0.0001). The expression of IL-22R1 (but not IL-20R1) was increased on monocytes from RA synovial fluid compared with monocytes from both RA and healthy control peripheral blood. The plasma concentrations of IL-20 and IL-24 were increased in rheumatoid factor and anti-citrullinated protein antibody positive compared with negative early RA patients (all P < 0.0001). Immune complexes stimulated the production of the IL-20R cytokines by monocytes/macrophages. Increased baseline plasma concentrations of IL-20 and IL-24 were associated with Sharp-van der Heijde score progression after 24 months (Spearman's rho = 0.19 and 0.26, both P < 0.05) in the early RA patients. The IL-22R1 was expressed by osteoclast precursors and in multinucleated osteoclasts. IL-20 and IL-24 increased the secretion of monocyte chemoattractant protein 1 by these cells.This study suggests that IL-20 and IL-24 link RA-associated autoantibodies with radiographic progression via the IL-22R1. Modulation of this axis holds promise as feasible anti-erosive treatment modalities in seropositive RA.

  View details for DOI 10.1186/s13075-016-0964-7

  View details for PubMedID 26968800

  View details for PubMedCentralID PMC4788924

• Contribution of Mast Cell-Derived Interleukin-1 beta to Uric Acid Crystal-Induced Acute Arthritis in Mice ARTHRITIS & RHEUMATOLOGY Reber, L. L., Marichal, T., Sokolove, J., Starkl, P., Gaudenzio, N., Iwakura, Y., Karasuyama, H., Schwartz, L. B., Robinson, W. H., Tsai, M., Galli, S. J. 2014; 66 (10): 2881-2891

  Abstract

  Gouty arthritis is caused by the precipitation of monosodium urate monohydrate (MSU) crystals in the joints. While it has been reported that mast cells (MCs) infiltrate gouty tophi, little is known about the actual roles of MCs during acute attacks of gout. This study was undertaken to assess the role of MCs in a mouse model of MSU crystal-induced acute arthritis.We assessed the effects of intraarticular (IA) injection of MSU crystals in various strains of mice with constitutive or inducible MC deficiency or in mice lacking interleukin-1β (IL-1β) or other elements of innate immunity. We also assessed the response to IA injection of MSU crystals in genetically MC-deficient mice after IA engraftment of wild-type or IL-1β(-/-) bone marrow-derived cultured MCs.MCs were found to augment acute tissue swelling following IA injection of MSU crystals in mice. IL-1β production by MCs contributed importantly to MSU crystal-induced tissue swelling, particularly during its early stages. Selective depletion of synovial MCs was able to diminish MSU crystal-induced acute inflammation in the joints.Our findings identify a previously unrecognized role of MCs and MC-derived IL-1β in the early stages of MSU crystal-induced acute arthritis in mice.

  View details for DOI 10.1002/art.38747

  View details for Web of Science ID 000342744300026

  View details for PubMedCentralID PMC4443497

• Smoking Status Is Associated with Inflammatory Cytokine Profile and Disease Activity: Decreased Inflammation and Disease Improvement with Smoking Cessation? Sokolove, J., Sayles, H., Wagner, C., Lahey, L. J., Thiele, G. M., Robinson, W. H., Reimold, A., Kerr, G. S., Cannon, G. W., Mikuls, T. R. WILEY-BLACKWELL. 2014: S146

  View details for Web of Science ID 000344384900347

• The Association of Fine Specificities of Anti-Citrullinated Protein Antibodies (ACPA) with disease Severity in African-Americans with RA. Danila, M. I., Reynolds, R., Wu, G., Tiwari, H., Robinson, W. H., Sokolove, J., Bridges, S., CLEAR Investigators WILEY-BLACKWELL. 2014: S192–S193

  View details for Web of Science ID 000344384900450

• Netosis Induced Histone Citrullination Facilitates Onset and Propagation of Rheumatoid Arthritis. Sohn, D., Onuma, K., Rhodes, C., Zhao, X., Gazitt, T., Shiao, R., Bober, J., Cheng, D., Lahey, L. J., Wong, H., van Eyk, J., Robinson, W. H., Sokolove, J. WILEY-BLACKWELL. 2014: S357–S358

  View details for Web of Science ID 000344384901359

• The Use of Multiplex Bead Array to Follow the Effect of Rituximab on IgG and IgA Serum Autoantibody Responses to Citrullinated Epitopes in Patients with Rheumatoid Arthritis. Cambridge, G., Lahey, L. J., Leandro, M. J., Robinson, W. H., Sokolove, J. WILEY-BLACKWELL. 2014: S193

  View details for Web of Science ID 000344384900451

• In a Periodontal Disease Cohort without RA, Indeterminate or Low-Positive Anti-CCP-2 Antibodies Are Associated with Multiple Distinct ACPA. Molitor, J. A., Michalowicz, B. S., Demmer, R. T., Buckner, J. H., Wener, M. H., Robinson, W. H. WILEY-BLACKWELL. 2014: S193–S194

  View details for Web of Science ID 000344384900452

• Vascular Calcifications on Hand and Wrist Radiographs Are Associated with Cardiovascular Risk Factors, Antigen-Specific Anti-Citrullinated Protein Antibodies, and Mortality in Rheumatoid Arthritis. Solow, E., Yu, F., Thiele, G. M., Sokolove, J., Robinson, W. H., Pruhs, Z. M., Michaud, K., Erickson, A. R., Sayles, H., Kerr, G. S., Gaffo, A. L., Caplan, L., Davis, L. A., Cannon, G. W., Reimold, A. M., Baker, J., Schwab, P., Anderson, D., Mikuls, T. R. WILEY-BLACKWELL. 2014: S370–S371

  View details for Web of Science ID 000344384901384

• Malondialdehyde-Acetaldehyde Adducts (MAA) and Anti-Malondialdehyde-Acetaldehyde Antibody in Rheumatoid Arthritis Thiele, G., Duryee, M. J., Anderson, D., Klassen, L. W., Mohring, S., Young, K., Benissan-Messan, D., Sayles, H., Sokolove, J., Robinson, W. H., Dell, J., Nicholas, A., Mikuls, T. R. WILEY-BLACKWELL. 2014: S646

  View details for Web of Science ID 000344384903094

• Antibodies to Citrullinated Clusterin, Filaggrin, Vimentin, and Fibrinogen Are Associated with Blood Pressure in First-Degree Relatives of Rheumatoid Arthritis Patients: The Studies of the Etiology of Rheumatoid Arthritis. Hughes-Austin, J. M., Gan, R. W., Deane, K. D., Gregersen, P. K., Weisman, M. H., Ix, J. H., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M. WILEY-BLACKWELL. 2014: S886–S887

  View details for Web of Science ID 000344384904169

• Associations of Toll-like Receptor (TLR)-4 Single Nucleotide Polymorphisms and Rheumatoid Arthritis Disease Progression. Davis, M., LeVan, T., Yu, F., Sayles, H., Sokolove, J., Robinson, W. H., Michaud, K., Thiele, G. M., Mikuls, T. R. WILEY-BLACKWELL. 2014: S1071

  View details for Web of Science ID 000344384905134

• A Dose Response Relationship Between Shared Epitope and ACPA Level: But Not All SE Alleles Are Created Equal. Sokolove, J., Lahey, L. J., Wagner, C., Smolik, I., Robinson, D. B., Ferucci, E. D., Newkirk, M., Fritzler, M., Robinson, W. H., El-Gabalawy, H. WILEY-BLACKWELL. 2014: S649

  View details for Web of Science ID 000344384903100

• Flip Deficiency in Dendritic Cells Promotes Spontaneous Arthritis Mediated By Reduced Treg and Increased Autoreactive CD4(+) t Cells. Huang, Q., Perlman, H. R., Birkett, R., Doyle, R. E., Fang, D., Haines, G., Robinson, W. H., Datta, S. K., Phee, H., Pope, R. M. WILEY-BLACKWELL. 2014: S1271

  View details for Web of Science ID 000344384906143

• Role of protein phosphatase magnesium-dependent 1A and anti-protein phosphatase magnesium-dependent 1A autoantibodies in ankylosing spondylitis. Arthritis & rheumatology (Hoboken, N.J.) Kim, Y. G., Sohn, D. H., Zhao, X., Sokolove, J., Lindstrom, T. M., Yoo, B., Lee, C. K., Reveille, J. D., Taurog, J. D., Robinson, W. H. 2014; 66 (10): 2793-2803

  Abstract

  Although ankylosing spondylitis (AS) is driven by immune-mediated processes, little is known about the presence and role of autoantibodies in this disease. This study was undertaken to investigate whether autoantibodies occur in and are involved in AS.We performed human protein microarray analysis of sera derived from patients with AS or other autoimmune disorders to identify autoantibodies associated specifically with AS, and identified autoantibody targeting of protein phosphatase magnesium-dependent 1A (PPM1A) in AS. We performed enzyme-linked immunosorbent assay (ELISA) analysis of sera from 2 independent AS cohorts to confirm autoantibody targeting of PPM1A, and to assess associations between levels of anti-PPM1A antibodies and AS disease severity or response to anti-tumor necrosis factor (anti-TNF) therapy (as measured by Bath AS Disease Activity Index [BASDAI] score). Levels of anti-PPM1A antibodies were also evaluated in sera from rats transgenic for HLA-B27 and human β2 -microglobulin. The expression of PPM1A was assessed by immunohistochemistry in synovial tissue samples from patients with AS, rheumatoid arthritis, or osteoarthritis. The role of PPM1A in osteoblast differentiation was investigated by gene knockdown and overexpression.AS was associated with autoantibody targeting of PPM1A, and levels of anti-PPM1A autoantibodies were significantly higher in patients with more advanced sacroiliitis and correlated positively with BASDAI score after treatment with anti-TNF agents. The levels of anti-PPM1A autoantibodies were also higher in the sera of transgenic rats that are prone to develop spondyloarthritis than in those that are not. PPM1A was expressed in AS synovial tissue, and PPM1A overexpression promoted osteoblast differentiation, whereas PPM1A knockdown suppressed it.Anti-PPM1A autoantibodies are present in AS, and our findings suggest that PPM1A may contribute to the pathogenic bone ankylosis characteristic of AS.

  View details for DOI 10.1002/art.38763

  View details for PubMedID 24980965

  View details for PubMedCentralID PMC4198528

• Barcode-enabled sequencing of plasmablast antibody repertoires in rheumatoid arthritis. Arthritis & rheumatology Tan, Y., Kongpachith, S., Blum, L. K., Ju, C., Lahey, L. J., Lu, D. R., Cai, X., Wagner, C. A., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 66 (10): 2706-2715

  Abstract

  A hallmark of rheumatoid arthritis (RA) is the production of autoantibodies, including anti-citrullinated protein antibodies (ACPAs). Nevertheless, the specific targets of these autoantibodies remain incompletely defined. During an immune response, B cells specific for the inciting antigen(s) are activated and differentiate into plasmablasts, which are released into the blood. We undertook this study to sequence the plasmablast antibody repertoire to define the targets of the active immune response in RA.We developed a novel DNA barcoding method to sequence the cognate heavy- and light-chain pairs of antibodies expressed by individual blood plasmablasts in RA. The method uses a universal 5' adapter that enables full-length sequencing of the antibodies' variable regions and recombinant expression of the paired antibody chains. The sequence data sets were bioinformatically analyzed to generate phylogenetic trees that identify clonal families of antibodies sharing heavy- and light-chain VJ sequences. Representative antibodies were expressed, and their binding properties were characterized using anti-cyclic citrullinated peptide 2 (anti-CCP-2) enzyme-linked immunosorbent assay (ELISA) and antigen microarrays.We used our sequencing method to generate phylogenetic trees representing the antibody repertoires of peripheral blood plasmablasts from 4 individuals with anti-CCP+ RA, and recombinantly expressed 14 antibodies that were either "singleton" antibodies or representative of clonal antibody families. Anti-CCP-2 ELISA identified 4 ACPAs, and antigen microarray analysis identified ACPAs that differentially targeted epitopes on α-enolase, citrullinated fibrinogen, and citrullinated histone H2B.Our data provide evidence that autoantibodies targeting α-enolase, citrullinated fibrinogen, and citrullinated histone H2B are produced by the ongoing activated B cell response in, and thus may contribute to the pathogenesis of, RA.

  View details for DOI 10.1002/art.38754

  View details for PubMedID 24965753

• Contribution of mast cell-derived interleukin-1ß to uric acid crystal-induced acute arthritis in mice. Arthritis & rheumatology Reber, L. L., Marichal, T., Sokolove, J., Starkl, P., Gaudenzio, N., Iwakura, Y., Karasuyama, H., Schwartz, L. B., Robinson, W. H., Tsai, M., Galli, S. J. 2014; 66 (10): 2881-2891

  Abstract

  Gouty arthritis is caused by the precipitation of monosodium urate monohydrate (MSU) crystals in the joints. While it has been reported that mast cells (MCs) infiltrate gouty tophi, little is known about the actual roles of MCs during acute attacks of gout. This study was undertaken to assess the role of MCs in a mouse model of MSU crystal-induced acute arthritis.We assessed the effects of intraarticular (IA) injection of MSU crystals in various strains of mice with constitutive or inducible MC deficiency or in mice lacking interleukin-1β (IL-1β) or other elements of innate immunity. We also assessed the response to IA injection of MSU crystals in genetically MC-deficient mice after IA engraftment of wild-type or IL-1β(-/-) bone marrow-derived cultured MCs.MCs were found to augment acute tissue swelling following IA injection of MSU crystals in mice. IL-1β production by MCs contributed importantly to MSU crystal-induced tissue swelling, particularly during its early stages. Selective depletion of synovial MCs was able to diminish MSU crystal-induced acute inflammation in the joints.Our findings identify a previously unrecognized role of MCs and MC-derived IL-1β in the early stages of MSU crystal-induced acute arthritis in mice.

  View details for DOI 10.1002/art.38747

  View details for PubMedID 24943488

• Stem Cell Growth Factor Expression in Rheumatoid Arthritis. Woo, Y., Baik, Y., Park, Y., Lee, S., Robinson, W. H., Song, J. WILEY-BLACKWELL. 2014: S642

  View details for Web of Science ID 000344384903081

• Role of Protein Phosphatase Magnesium-Dependent 1A and Anti-Protein Phosphatase Magnesium-Dependent 1A Autoantibodies in Ankylosing Spondylitis ARTHRITIS & RHEUMATOLOGY Kim, Y., Sohn, D. H., Zhao, X., Sokolove, J., Lindstrom, T. M., Yoo, B., Lee, C., Reveille, J. D., Taurog, J. D., Robinson, W. H. 2014; 66 (10): 2793-2803

  Abstract

  Although ankylosing spondylitis (AS) is driven by immune-mediated processes, little is known about the presence and role of autoantibodies in this disease. This study was undertaken to investigate whether autoantibodies occur in and are involved in AS.We performed human protein microarray analysis of sera derived from patients with AS or other autoimmune disorders to identify autoantibodies associated specifically with AS, and identified autoantibody targeting of protein phosphatase magnesium-dependent 1A (PPM1A) in AS. We performed enzyme-linked immunosorbent assay (ELISA) analysis of sera from 2 independent AS cohorts to confirm autoantibody targeting of PPM1A, and to assess associations between levels of anti-PPM1A antibodies and AS disease severity or response to anti-tumor necrosis factor (anti-TNF) therapy (as measured by Bath AS Disease Activity Index [BASDAI] score). Levels of anti-PPM1A antibodies were also evaluated in sera from rats transgenic for HLA-B27 and human β2 -microglobulin. The expression of PPM1A was assessed by immunohistochemistry in synovial tissue samples from patients with AS, rheumatoid arthritis, or osteoarthritis. The role of PPM1A in osteoblast differentiation was investigated by gene knockdown and overexpression.AS was associated with autoantibody targeting of PPM1A, and levels of anti-PPM1A autoantibodies were significantly higher in patients with more advanced sacroiliitis and correlated positively with BASDAI score after treatment with anti-TNF agents. The levels of anti-PPM1A autoantibodies were also higher in the sera of transgenic rats that are prone to develop spondyloarthritis than in those that are not. PPM1A was expressed in AS synovial tissue, and PPM1A overexpression promoted osteoblast differentiation, whereas PPM1A knockdown suppressed it.Anti-PPM1A autoantibodies are present in AS, and our findings suggest that PPM1A may contribute to the pathogenic bone ankylosis characteristic of AS.

  View details for DOI 10.1002/art.38763

  View details for Web of Science ID 000342744300016

  View details for PubMedCentralID PMC4198528

• Reply: To PMID 24497204. Arthritis & rheumatology Shelef, M. A., Sokolove, J., Robinson, W. H., Huttenlocher, A. 2014; 66 (9): 2644-2645

  View details for DOI 10.1002/art.38722

  View details for PubMedID 24891318

• Street-experienced peripheral B cells traffic to the brain. Science translational medicine Lu, D. R., Robinson, W. H. 2014; 6 (248): 248fs31-?

  Abstract

  Deep sequencing of immunoglobulin repertoires in multiple sclerosis patients reveals dysregulation of peripheral B cell immunity (Palanichamy et al. and Stern et al., this issue).

  View details for DOI 10.1126/scitranslmed.3009919

  View details for PubMedID 25100737

• Quantitative Radiologic Imaging Techniques for Articular Cartilage Composition: Toward Early Diagnosis and Development of Disease-Modifying Therapeutics for Osteoarthritis ARTHRITIS CARE & RESEARCH Oei, E. H., van Tiel, J., Robinson, W. H., Gold, G. E. 2014; 66 (8): 1129-1141

  View details for DOI 10.1002/acr.22316

  View details for PubMedID 24578345

• Optical imaging of articular cartilage degeneration using near-infrared dipicolylamine probes. Biomaterials Hu, X., Wang, Q., Liu, Y., Liu, H., Qin, C., Cheng, K., Robinson, W., Gray, B. D., Pak, K. Y., Yu, A., Cheng, Z. 2014; 35 (26): 7511-7521

  Abstract

  Articular cartilage is the hydrated tissue that lines the ends of long bones in load bearing joints and provides joints with a smooth, nearly frictionless gliding surface. However, the deterioration of articular cartilage occurs in the early stages of osteoarthritis (OA) and is clinically and radiographically silent. Here two cationic near infrared fluorescent (NIRF) dipicolylamine (DPA) probes, Cy5-DPA-Zn and Cy7-DPA-Zn, were prepared for cartilage degeneration imaging and OA early detection through binding to the anionic glycosaminoglycans (GAGs). The feasibility of NIRF dye labeled DPA-Zn probes for cartilage degeneration imaging was examined ex vivo and in vivo. The ex vivo studies showed that Cy5-DPA-Zn and Cy7-DPA-Zn not only showed the high uptake and electrostatic attractive binding to cartilage, but also sensitively reflected the change of GAGs contents. In vivo imaging study further indicated that Cy5-DPA-Zn demonstrated higher uptake and retention in young mice (high GAGs) than old mice (low GAGs) when administrated via local injection in mouse knee joints. More importantly, Cy5-DPA-Zn showed dramatic higher signals in sham joint (high GAGs) than OA side (low GAGs), through sensitive reflecting the change of GAGs in the surgical induced OA models. In summary, Cy5-DPA-Zn provides promising visual detection for early cartilage pathological degeneration in living subjects.

  View details for DOI 10.1016/j.biomaterials.2014.05.042

  View details for PubMedID 24912814

• Association of fine specificity and repertoire expansion of anticitrullinated peptide antibodies with rheumatoid arthritis associated interstitial lung disease ANNALS OF THE RHEUMATIC DISEASES Giles, J. T., Danoff, S. K., Sokolove, J., Wagner, C. A., Winchester, R., Pappas, D. A., Siegelman, S., Connors, G., Robinson, W. H., Bathon, J. M. 2014; 73 (8): 1487-1494

  Abstract

  Interstitial lung disease (ILD) is associated with high morbidity and mortality in rheumatoid arthritis (RA). Citrullinated proteins are observed in RA lung tissues; however, the association of specific anticitrullinated peptide antibodies (ACPA) with ILD in RA is unknown.RA patients underwent multidetector CT (MDCT) of the chest, from which ILD features and a semiquantitative ILD Score (ILDS; range 0-32) were assessed. Anti-CCP (CCP2) and levels of a panel of antibodies against 17 citrullinated and four non-citrullinated peptides were assessed from concurrent serum samples using a custom Bio-Plex bead array. High level ACPA was defined as ≥the group 75th percentile.Among the 177 RA patients studied, median levels of CCP2 and all specific ACPAs were 46-273% higher among RA patients with versus those without ILD (all p values <0.05), and higher levels correlated with higher ILDS. In contrast, levels of non-citrullinated protein antibodies were not higher in those with ILD. RA patients had a median of 2 high level ACPA reactivities (range 0-16), with each high level ACPA associated, on average, with a 0.10 unit higher ILDS (p=0.001). This association remained significant after adjusting for characteristics associated with ILD (age, gender, current and former smoking, Disease Activity Score for 28 joints, current prednisone and leflunomide use). More high level ACPA were observed in those with versus without pulmonary function restriction or impaired diffusion.Our findings of a broader ACPA repertoire in RA ILD suggest a possible role for ACPA in the pathogenesis of ILD.

  View details for DOI 10.1136/annrheumdis-2012-203160

  View details for PubMedID 23716070

• Peptidylarginine Deiminase 4 Contributes to Tumor Necrosis Factor alpha-Induced Inflammatory Arthritis ARTHRITIS & RHEUMATOLOGY Shelef, M. A., Sokolove, J., Lahey, L. J., Wagner, C. A., Sackmann, E. K., Warner, T. F., Wang, Y., Beebe, D. J., Robinson, W. H., Huttenlocher, A. 2014; 66 (6): 1482-1491

  Abstract

  Peptidylarginine deiminase 4 (PAD4) is a citrullinating enzyme that has multiple associations with inflammation. In rheumatoid arthritis, PAD4 and protein citrullination are increased in inflamed joints, and anti-citrullinated protein antibodies (ACPAs) form against citrullinated antigens are formed. ACPA immune complexes can deposit in the joint and induce the production of tumor necrosis factor α (TNFα), a critical inflammatory cytokine in the pathogenesis of rheumatoid arthritis. Further, in other settings, TNFα has been shown to induce PAD4 activity and modulate antibody formation. We undertook this study to investigate whether TNFα and PAD4 may synergistically exacerbate autoantibody production and inflammatory arthritis.To determine whether TNFα and PAD4 augment autoantibody production and inflammatory arthritis, we first used a multiplex assay to determine whether mice with chronic inflammatory arthritis due to overexpression of TNFα develop autoantibodies against native and citrullinated antigens. With TNF(+) PAD4(+/+) and TNF(+) PAD4(-/-) mice, we then compared serum autoantibody levels by multiplex array, lymphocyte activation by flow cytometry, total serum IgG levels by enzyme-linked immunosorbent assay, arthritis by clinical and histologic scoring, and systemic inflammation using microfluidic devices.TNFα-overexpressing mice had increased levels of autoantibodies reactive against native and citrullinated antigens. PAD4(-/-) mice with TNFα-induced arthritis had lower levels of autoantibodies reactive against native and citrullinated antigens, decreased T cell activation and total IgG levels, and reduced inflammation and arthritis compared to PAD4(+/+) TNFα-overexpressing mice.PAD4 mediates autoantibody production and inflammatory arthritis downstream of TNFα.

  View details for DOI 10.1002/art.38393

  View details for Web of Science ID 000337366200010

  View details for PubMedCentralID PMC4148484

• Periodontitis and Porphyromonas gingivalis in Patients With Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Mikuls, T. R., Payne, J. B., Yu, F., Thiele, G. M., Reynolds, R. J., Cannon, G. W., Markt, J., McGowan, D., Kerr, G. S., Redman, R. S., Reimold, A., Griffiths, G., Beatty, M., Gonzalez, S. M., Bergman, D. A., Hamilton, B. C., Erickson, A. R., Sokolove, J., Robinson, W. H., Walker, C., Chandad, F., O'Dell, J. R. 2014; 66 (5): 1090-1100

  Abstract

  To examine the degree to which shared risk factors explain the relationship of periodontitis (PD) to rheumatoid arthritis (RA) and to determine the associations of PD and Porphyromonas gingivalis with pathologic and clinical features of RA.Patients with RA (n = 287) and patients with osteoarthritis as disease controls (n = 330) underwent a standardized periodontal examination. The HLA-DRB1 status of all participants was imputed using single-nucleotide polymorphisms from the extended major histocompatibility complex. Circulating anti-P gingivalis antibodies were measured using an enzyme-linked immunosorbent assay, and subgingival plaque was assessed for the presence of P gingivalis using polymerase chain reaction (PCR). Associations of PD with RA were examined using multivariable regression.Presence of PD was more common in patients with RA and patients with anti-citrullinated protein antibody (ACPA)-positive RA (n = 240; determined using the anti-cyclic citrullinated peptide 2 [anti-CCP-2] test) than in controls (35% and 37%, respectively, versus 26%; P = 0.022 and P = 0.006, respectively). There were no differences between RA patients and controls in the levels of anti-P gingivalis or the frequency of P gingivalis positivity by PCR. The anti-P gingivalis findings showed a weak, but statistically significant, association with the findings for both anti-CCP-2 (r = 0.14, P = 0.022) and rheumatoid factor (RF) (r = 0.19, P = 0.001). Presence of PD was associated with increased swollen joint counts (P = 0.004), greater disease activity according to the 28-joint Disease Activity Score using C-reactive protein level (P = 0.045), and higher total Sharp scores of radiographic damage (P = 0.015), as well as with the presence and levels of anti-CCP-2 (P = 0.011) and RF (P < 0.001). The expression levels of select ACPAs (including antibodies to citrullinated filaggrin) were higher in patients with subgingival P gingivalis and in those with higher levels of anti-P gingivalis antibodies, irrespective of smoking status. Associations of PD with established seropositive RA were independent of all covariates examined, including evidence of P gingivalis infection.Both PD and P gingivalis appear to shape the autoreactivity of RA. In addition, these results demonstrate an independent relationship between PD and established seropositive RA.

  View details for DOI 10.1002/art.38348

  View details for Web of Science ID 000337363400006

  View details for PubMedCentralID PMC4115329

• Identifying functional anti-Staphylococcus aureus antibodies by sequencing antibody repertoires of patient plasmablasts CLINICAL IMMUNOLOGY Lu, D. R., Tan, Y., Kongpachith, S., Cai, X., Stein, E. A., Lindstrom, T. M., Sokolove, J., Robinson, W. H. 2014; 152 (1-2): 77-89

  Abstract

  Infection by Staphylococcus aureus is on the rise, and there is a need for a better understanding of host immune responses that combat S. aureus. Here we use DNA barcoding to enable deep sequencing of the paired heavy- and light-chain immunoglobulin genes expressed by individual plasmablasts derived from S. aureus-infected humans. Bioinformatic analysis of the antibody repertoires revealed clonal families of heavy-chain sequences and enabled rational selection of antibodies for recombinant expression. Of the ten recombinant antibodies produced, seven bound to S. aureus, of which four promoted opsonophagocytosis of S. aureus. Five of the antibodies bound to known S. aureus cell-surface antigens, including fibronectin-binding protein A. Fibronectin-binding protein A-specific antibodies were isolated from two independent S. aureus-infected patients and mediated neutrophil killing of S. aureus in in vitro assays. Thus, our DNA barcoding approach enabled efficient identification of antibodies involved in protective host antibody responses against S. aureus.

  View details for DOI 10.1016/j.clim.2014.02.010

  View details for PubMedID 24589749

• Rheumatoid Factor as a Potentiator of Anti-Citrullinated Protein Antibody-Mediated Inflammation in Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Sokolove, J., Johnson, D. S., Lahey, L. J., Wagner, C. A., Cheng, D., Thiele, G. M., Michaud, K., Sayles, H., Reimold, A. M., Caplan, L., Cannon, G. W., Kerr, G., Mikuls, T. R., Robinson, W. H. 2014; 66 (4): 813-821

  Abstract

  The co-occurrence of rheumatoid factor (RF) and anti-citrullinated protein antibody (ACPA) positivity in rheumatoid arthritis (RA) is well described. However, the mechanisms underlying the potential interaction between these 2 distinct autoantibodies have not been well defined. The aim of this study was to evaluate the epidemiologic and molecular interaction of ACPAs and RF and its association with both disease activity and measures of RA-associated inflammation.In a cohort of 1,488 US veterans with RA, measures of disease activity and serum levels of cytokines and multiplex ACPAs were compared between the following groups of patients: double-negative (anti-cyclic citrullinated peptide [anti-CCP]-/RF-), anti-CCP+/RF-, anti-CCP-/RF+, or double-positive (anti-CCP+/RF+). Additional studies were performed using an in vitro immune complex (IC) stimulation assay in which macrophages were incubated with ACPA ICs in the presence or absence of monoclonal IgM-RF, and tumor necrosis factor α production measured as a readout of macrophage activation.Compared with the double-negative subgroup (as well as each single-positive subgroup), the double-positive subgroup exhibited higher disease activity as well as higher levels of C-reactive protein and inflammatory cytokines (all P < 0.001). In vitro stimulation of macrophages by ACPA ICs increased cytokine production, and the addition of monoclonal IgM-RF significantly increased macrophage tumor necrosis factor α production (P = 0.003 versus ACPA ICs alone).The combined presence of ACPAs and IgM-RF mediates increased proinflammatory cytokine production in vitro and is associated with increased systemic inflammation and disease activity in RA. Our data suggest that IgM-RF enhances the capacity of ACPA ICs to stimulate macrophage cytokine production, thereby providing a mechanistic link by which RF enhances the pathogenicity of ACPA ICs in RA.

  View details for DOI 10.1002/art.38307

  View details for PubMedID 24757134

• Rheumatoid Arthritis in the Womens Health Initiative: Methods and Baseline Evaluation AMERICAN JOURNAL OF EPIDEMIOLOGY Kuller, L. H., Mackey, R. H., Walitt, B. T., Deane, K. D., Holers, V. M., Robinson, W. H., Sokolove, J., Chang, Y., Moreland, L. W. 2014; 179 (7): 917-926

  Abstract

  Second-generation assays for anti-cyclic citrullinated peptide (anti-CCP), a highly sensitive and specific marker for rheumatoid arthritis (RA), have redefined the epidemiology of RA. In the Women's Health Initiative (WHI) RA study (2009-2011), we evaluated the prevalence of anti-CCP positivity among 15,691 (10.2% of 161,808) WHI participants aged 50-79 years who reported RA. Using stored baseline specimens, we measured serum anti-CCP, rheumatoid factor (RF), and antinuclear antibody in a defined sample of 9,988 of black, white, and Hispanic women. In a subset of women, we measured plasma cytokine levels and number of copies of the human leukocyte antigen (HLA)-DRB1 (HLA-DRB1) shared epitope in DNA by means of Luminex polymerase chain reaction typing (Luminex Corporation, Austin, Texas). We validated classification of probable clinical RA in 2 clinics as anti-CCP positivity or self-reported validated use of disease-modifying antirheumatic drugs (DMARDs). The prevalence of anti-CCP positivity was 8.1%, and the prevalence of RF positivity was approximately 16.0%. DMARD use including prednisone was reported by 1,140 (11.4%) participants (841 excluding prednisone) but by 57.5% of anti-CCP-positive women. The prevalence of 2 shared epitopes was also much higher for anti-CCP-positive women (18.2%, as opposed to only 5.5% for women with anti-CCP-negative DMARD-positive RA and 6.6% for anti-CCP-negative, RF-negative DMARD nonusers). Median cytokine levels were much higher for anti-CCP-positive/RF-positive women. Women with anti-CCP-positive RA and anti-CCP-negative RA had different characteristics with regard to HLA shared epitope, cigarette smoking, and inflammation (cytokines).

  View details for DOI 10.1093/aje/kwu003

  View details for Web of Science ID 000333247200013

  View details for PubMedID 24569640

  View details for PubMedCentralID PMC3969537

• Determinants of Mortality Among Postmenopausal Women in the Women's Health Initiative Who Report Rheumatoid Arthritis ARTHRITIS & RHEUMATOLOGY Kuller, L. H., Mackey, R. H., Walitt, B. T., Deane, K. D., Holers, V. M., Robinson, W. H., Sokolove, J., Chang, Y., Liu, S., Parks, C. G., Wright, N. C., Moreland, L. W. 2014; 66 (3): 497-507

  Abstract

  Rheumatoid arthritis (RA) patients have an increased risk of cardiovascular disease (CVD) and mortality. We measured anti-cyclic citrullinated peptide (anti-CCP) antibody levels and determined use of disease-modifying antirheumatic drugs (DMARDs) among women in the Women's Health Initiative (WHI). Using these data, we undertook this study to assess total mortality over 10 years of followup among white, black, or Hispanic women with self-reported RA in the WHI.Using stored baseline serum, we measured anti-CCP, rheumatoid factor (RF), and antinuclear antibodies (ANAs) in 9,988 women who reported having RA. Based on a previous chart review study, probable RA was defined as either self-reported RA and anti-CCP positivity, or anti-CCP negativity and DMARD use. Cox proportional hazards regression was used to model the relationship of self-reported RA, DMARD exposure, and anti-CCP positivity to total mortality, using followup data through April 2009.At baseline, the mean age was 62.8 years; 24.5% of subjects were black and 10% were Hispanic. Prevalence of anti-CCP positivity was 8.1% (n = 812), and 217 women were anti-CCP negative but had reported use of DMARDs; therefore, 1,029 women (of 9,988) were classified as having probable RA, and 8,958 were classified as unlikely to have RA (with data on DMARD use missing for 1 subject). Age-adjusted mortality rates were ∼2-fold higher for anti-CCP-positive women, with 20.2 deaths per 1,000 person-years, as compared to 11.4 deaths per 1,000 person-years among anti-CCP-negative women with self-reported RA who never used DMARDs. Among women who did not report any arthritis at baseline, we found 8.3 deaths per 1,000 person-years. The increased risk among anti-CCP-positive women with RA was not explained by age, RF positivity, ANA positivity, or DMARD use.Anti-CCP-positive RA was associated with substantial excess mortality among postmenopausal women in the WHI. This result was not explained by the risk factors we measured.

  View details for DOI 10.1002/art.38268

  View details for Web of Science ID 000337359400004

  View details for PubMedID 24574208

• MONOCLONAL ANTIBODIES FROM CD19(+) SYNOVIAL B CELLS OF RA PATIENTS WITH TERTIARY LYMPHOID STRUCTURES DISPLAY A STRONG IMMUNOREACTIVITY TOWARDS CITRULLINATED HISTONES FROM NEUTROPHILS NETS Corsiero, E., Carlotti, E., Wardemann, H., Robinson, W., Migliorini, P., Pratesi, F., Pitzalis, C., Bombardieri, M. BMJ PUBLISHING GROUP. 2014: A13

  View details for DOI 10.1136/annrheumdis-2013-205124.30

  View details for Web of Science ID 000346248400031

• Brief report: carboxypeptidase B serves as a protective mediator in osteoarthritis. Arthritis & rheumatology (Hoboken, N.J.) Lepus, C. M., Song, J. J., Wang, Q., Wagner, C. A., Lindstrom, T. M., Chu, C. R., Sokolove, J., Leung, L. L., Robinson, W. H. 2014; 66 (1): 101-106

  Abstract

  We previously demonstrated that carboxypeptidase B (CPB) protects against joint erosion in rheumatoid arthritis by inactivating complement component C5a. We also found that levels of CPB are abnormally high in the synovial fluid of individuals with another joint disease, osteoarthritis (OA). We undertook this study to investigate whether CPB plays a role in the pathogenesis of OA.We compared the development of OA in CPB-deficient (Cpb2(-/-) ) mice and wild-type mice by subjecting them to medial meniscectomy and histologically assessing cartilage damage, osteophyte formation, and synovitis in the stifle joints 4 months later. We measured levels of proCPB, proinflammatory cytokines, and complement components in synovial fluid samples from patients with symptomatic and radiographic knee OA. Finally, we used enzyme-linked immunosorbent assay, flow cytometry, and hemolytic assays to assess the effect of CPB on formation of membrane attack complex (MAC)-a complement effector critical to OA pathogenesis.Cpb2(-/-) mice developed dramatically greater cartilage damage than did wild-type mice (P < 0.01) and had a greater number of osteophytes (P < 0.05) and a greater degree of synovitis (P < 0.05). In synovial fluid samples from OA patients, high levels of proCPB were associated with high levels of proinflammatory cytokines and complement components, and levels of proCPB correlated positively with those of MAC. In in vitro complement activation assays, activated CPB suppressed the formation of MAC as well as MAC-induced hemolysis.Our data suggest that CPB protects against inflammatory destruction of the joints in OA, at least in part by inhibiting complement activation.

  View details for DOI 10.1002/art.38213

  View details for PubMedID 24449579

• Decreased plasma levels of soluble CD18 link leukocyte infiltration with disease activity in spondyloarthritis. Arthritis research & therapy Kragstrup, T. W., Jalilian, B., Hvid, M., Kjærgaard, A., Østgård, R., Schiøttz-Christensen, B., Jurik, A. G., Robinson, W. H., Vorup-Jensen, T., Deleuran, B. 2014; 16 (1): R42-?

  Abstract

  Spondyloarthritis (SpA) comprises a group of diseases often associated with HLA-B27 and characterized by inflammation of the entheses and joints of the axial skeleton. The inflammatory process in SpA is presumably driven by innate immune cells but is still poorly understood. Thus, new tools for monitoring and treating inflammation are needed. The family of CD18 integrins is pivotal in guiding leukocytes to sites of inflammation, and CD18 hypomorphic mice develop a disease resembling SpA. Previously, we demonstrated that altered soluble CD18 (sCD18) complexes in the blood and synovial fluid of patients with arthritis have anti-inflammatory functions. Here, we study the mechanisms for these alterations and their association with SpA disease activity.Plasma levels of sCD18 in a study population with 84 patients with SpA and matched healthy controls were analyzed with a time-resolved immunoflourometric assay (TRIFMA). Binding of sCD18 to endothelial cells and fibroblast-like synoviocytes (FLSs) was studied with confocal microscopy. Shedding of CD18 from peripheral blood mononuclear cells (PBMCs) was studied with flow cytometry and TRIFMA.Plasma levels of sCD18 were decreased in patients with SpA compared with healthy volunteers (P <0.001), and the lowest levels were in the HLA-B27-positive subgroup (P <0.05). In a multiple regression model, the sCD18 levels exhibited an inverse correlation with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) (P <0.05), the level of morning stiffness (P <0.05), the Bath Ankylosing Spondilitis Metrology Index (P <0.05), the physician global assessment score (P <0.01), and the sacroiliac magnetic resonance imaging activity score (P <0.05). The mechanisms for these changes could be simulated in vitro. First, sCD18 in plasma adhered to inflammation-induced intercellular adhesion molecule 1 (ICAM-1) on endothelial cells and FLS, indicating increased consumption. Second, CD18 shedding from SpA PBMCs correlated inversely with the BASDAI (P <0.05), suggesting insufficient generation. CD18 was shed primarily from intermediate CD14⁺⁺ CD16⁺ monocytes, supporting the view that alterations in innate immunity can regulate the inflammatory processes in SpA.Taken together, the failure of patients with SpA to maintain adequate sCD18 levels may reflect insufficient CD18 shedding from monocytes to counterbalance the capture of sCD18 complexes to inflammation-induced ICAM-1. This could increase the availability of ICAM-1 molecules on the endothelium and in the synovium, facilitating leukocyte migration to the entheses and joints and aggregating disease activity.

  View details for DOI 10.1186/ar4471

  View details for PubMedID 24490631

  View details for PubMedCentralID PMC3978678

• Decreased plasma levels of soluble CD18 link leukocyte infiltration with disease activity in spondyloarthritis ARTHRITIS RESEARCH & THERAPY Kragstrup, T. W., Jalilian, B., Hvid, M., Kjaergaard, A., Ostgard, R., Schiottz-Christensen, B., Jurik, A. G., Robinson, W. H., Vorup-Jensen, T., Deleuran, B. 2014; 16 (1)

  Abstract

  Spondyloarthritis (SpA) comprises a group of diseases often associated with HLA-B27 and characterized by inflammation of the entheses and joints of the axial skeleton. The inflammatory process in SpA is presumably driven by innate immune cells but is still poorly understood. Thus, new tools for monitoring and treating inflammation are needed. The family of CD18 integrins is pivotal in guiding leukocytes to sites of inflammation, and CD18 hypomorphic mice develop a disease resembling SpA. Previously, we demonstrated that altered soluble CD18 (sCD18) complexes in the blood and synovial fluid of patients with arthritis have anti-inflammatory functions. Here, we study the mechanisms for these alterations and their association with SpA disease activity.Plasma levels of sCD18 in a study population with 84 patients with SpA and matched healthy controls were analyzed with a time-resolved immunoflourometric assay (TRIFMA). Binding of sCD18 to endothelial cells and fibroblast-like synoviocytes (FLSs) was studied with confocal microscopy. Shedding of CD18 from peripheral blood mononuclear cells (PBMCs) was studied with flow cytometry and TRIFMA.Plasma levels of sCD18 were decreased in patients with SpA compared with healthy volunteers (P <0.001), and the lowest levels were in the HLA-B27-positive subgroup (P <0.05). In a multiple regression model, the sCD18 levels exhibited an inverse correlation with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) (P <0.05), the level of morning stiffness (P <0.05), the Bath Ankylosing Spondilitis Metrology Index (P <0.05), the physician global assessment score (P <0.01), and the sacroiliac magnetic resonance imaging activity score (P <0.05). The mechanisms for these changes could be simulated in vitro. First, sCD18 in plasma adhered to inflammation-induced intercellular adhesion molecule 1 (ICAM-1) on endothelial cells and FLS, indicating increased consumption. Second, CD18 shedding from SpA PBMCs correlated inversely with the BASDAI (P <0.05), suggesting insufficient generation. CD18 was shed primarily from intermediate CD14⁺⁺ CD16⁺ monocytes, supporting the view that alterations in innate immunity can regulate the inflammatory processes in SpA.Taken together, the failure of patients with SpA to maintain adequate sCD18 levels may reflect insufficient CD18 shedding from monocytes to counterbalance the capture of sCD18 complexes to inflammation-induced ICAM-1. This could increase the availability of ICAM-1 molecules on the endothelium and in the synovium, facilitating leukocyte migration to the entheses and joints and aggregating disease activity.

  View details for DOI 10.1186/ar4471

  View details for Web of Science ID 000335148700041

  View details for PubMedCentralID PMC3978678

• Oral and topical boswellic acid attenuates mouse osteoarthritis OSTEOARTHRITIS AND CARTILAGE Wang, Q., Pan, X., Wong, H. H., Wagner, C. A., Lahey, L. J., Robinson, W. H., Sokolove, J. 2014; 22 (1): 128-132

  Abstract

  Boswellic acid is a plant-derived molecule with putative anti-inflammatory effects. This study was performed to determine whether oral or topical administration of boswellic acid can attenuate joint damage in a mouse model of osteoarthritis (OA).Levels of boswellic acid were measured in the blood and synovium of mice treated with oral or topical boswellic acid. OA was generated by surgical destabilization of the medial meniscus (DMM). Therapy with oral or topical boswellic acid was initiated one day after surgery and continued for 12 weeks, when knees were harvested and scored histologically for degree of cartilage loss, osteophyte formation, and synovitis. Microdissected OA synovium was stimulated with IL-1β or lipopolysaccharide (LPS) in the presence or absence of boswellic acid and cytokine production by quantitative polymerase chain reaction (PCR) or multiplex enzyme linked immunoabsorbant assay (ELISA).Topical treatment resulted in synovial concentrations of boswellic acid 2-6-fold higher than that measured in plasma. Cartilage loss was significantly reduced in mice treated with oral or topical boswellic acid compared with vehicle control (P < 0.01 for both oral and topical therapies). Likewise, treatment with either oral boswellic acid or boswellic acid ointment reduced of synovitis (P = 0.006 and 0.025, respectively) and osteophyte formation (P = 0.009 and 0.030, respectively). In vitro, boswellic acid was able to inhibit IL-1β and TLR4 mediated induction of several inflammatory mediators from OA synovial explant tissue.Significant synovial concentration and therapeutic efficacy can be achieved with topical boswellic acid treatment. These findings suggest that boswellic acid has potential as a disease-modifying agent in OA.

  View details for DOI 10.1016/j.joca.2013.10.012

  View details for Web of Science ID 000330422000016

  View details for PubMedID 24185109

  View details for PubMedCentralID PMC3992997

• Serum inflammatory mediators as markers of human lyme disease activity. PloS one Soloski, M. J., Crowder, L. A., Lahey, L. J., Wagner, C. A., Robinson, W. H., Aucott, J. N. 2014; 9 (4)

  Abstract

  Chemokines and cytokines are key signaling molecules that orchestrate the trafficking of immune cells, direct them to sites of tissue injury and inflammation and modulate their states of activation and effector cell function. We have measured, using a multiplex-based approach, the levels of 58 immune mediators and 7 acute phase markers in sera derived from of a cohort of patients diagnosed with acute Lyme disease and matched controls. This analysis identified a cytokine signature associated with the early stages of infection and allowed us to identify two subsets (mediator-high and mediator-low) of acute Lyme patients with distinct cytokine signatures that also differed significantly (p<0.0005) in symptom presentation. In particular, the T cell chemokines CXCL9 (MIG), CXCL10 (IP-10) and CCL19 (MIP3B) were coordinately increased in the mediator-high group and levels of these chemokines could be associated with seroconversion status and elevated liver function tests (p = 0.027 and p = 0.021 respectively). There was also upregulation of acute phase proteins including CRP and serum amyloid A. Consistent with the role of CXCL9/CXCL10 in attracting immune cells to the site of infection, CXCR3+ CD4 T cells are reduced in the blood of early acute Lyme disease (p = 0.01) and the decrease correlates with chemokine levels (p = 0.0375). The levels of CXCL9/10 did not relate to the size or number of skin lesions but elevated levels of serum CXCL9/CXCL10 were associated with elevated liver enzymes levels. Collectively these results indicate that the levels of serum chemokines and the levels of expression of their respective chemokine receptors on T cell subsets may prove to be informative biomarkers for Lyme disease and related to specific disease manifestations.

  View details for DOI 10.1371/journal.pone.0093243

  View details for PubMedID 24740099

• Carboxypeptidase B Serves as a Protective Mediator in Osteoarthritis ARTHRITIS & RHEUMATOLOGY Lepus, C. M., Song, J. J., Wang, Q., Wagner, C. A., Lindstrom, T. M., Chu, C. R., Sokolove, J., Leung, L. L., Robinson, W. H. 2014; 66 (1): 101-106

  Abstract

  We previously demonstrated that carboxypeptidase B (CPB) protects against joint erosion in rheumatoid arthritis by inactivating complement component C5a. We also found that levels of CPB are abnormally high in the synovial fluid of individuals with another joint disease, osteoarthritis (OA). We undertook this study to investigate whether CPB plays a role in the pathogenesis of OA.We compared the development of OA in CPB-deficient (Cpb2(-/-) ) mice and wild-type mice by subjecting them to medial meniscectomy and histologically assessing cartilage damage, osteophyte formation, and synovitis in the stifle joints 4 months later. We measured levels of proCPB, proinflammatory cytokines, and complement components in synovial fluid samples from patients with symptomatic and radiographic knee OA. Finally, we used enzyme-linked immunosorbent assay, flow cytometry, and hemolytic assays to assess the effect of CPB on formation of membrane attack complex (MAC)-a complement effector critical to OA pathogenesis.Cpb2(-/-) mice developed dramatically greater cartilage damage than did wild-type mice (P < 0.01) and had a greater number of osteophytes (P < 0.05) and a greater degree of synovitis (P < 0.05). In synovial fluid samples from OA patients, high levels of proCPB were associated with high levels of proinflammatory cytokines and complement components, and levels of proCPB correlated positively with those of MAC. In in vitro complement activation assays, activated CPB suppressed the formation of MAC as well as MAC-induced hemolysis.Our data suggest that CPB protects against inflammatory destruction of the joints in OA, at least in part by inhibiting complement activation.

  View details for DOI 10.1002/art.38213

  View details for Web of Science ID 000337356300014

• Clinical optimization of antigen specific modulation of type 1 diabetes with the plasmid DNA platform CLINICAL IMMUNOLOGY Gottlieb, P., Utz, P. J., Robinson, W., Steinman, L. 2013; 149 (3): 297-306

  Abstract

  Some clinical trials in humans have aimed at modulation of type 1 diabetes (T1D) via alteration of the immune response to putative islet cell antigens, particularly proinsulin and insulin, glutamic acid decarboxylase and the peptide, DiaPep 277, derived from heat shock protein 60. The focus here is on development of a specially engineered DNA plasmid encoding proinsulin to treat T1D. The plasmid is engineered to turn off adaptive immunity to proinsulin. This approach yielded exciting results in a randomized placebo controlled trial in 80 adult patients with T1D. The implications of this trial are explored in regards to the potential for sparing inflammation in islets and thus allowing the functioning beta cells to recover and produce more insulin. Strategies to further strengthen the effects seen thus far with the tolerizing DNA plasmid to proinsulin will be elucidated. The DNA platform affords an opportunity for easy modifications. In addition standard exploration of dose levels, route of administration and frequency of dose are practical. Optimization of the effects seen to date on C-peptide and on depletion of proinsulin specific CD8 T cells are feasible, with expected concomitant improvement in other parameters like hemoglobin A1c and reduction in insulin usage. T1D is one of the few autoimmune conditions where antigen specific therapy can be achieved, provided the approach is tested intelligently. Tolerizing DNA vaccines to proinsulin and other islet cell autoantigens is a worthy pursuit to potentially treat, prevent and to perhaps even 'cure' or 'prevent' type 1 diabetes.

  View details for DOI 10.1016/j.clim.2013.08.010

  View details for Web of Science ID 000328874700005

  View details for PubMedID 24094739

• Protein microarray analysis reveals BAFF-binding autoantibodies in systemic lupus erythematosus JOURNAL OF CLINICAL INVESTIGATION Price, J. V., Haddon, D. J., Kemmer, D., Delepine, G., Mandelbaum, G., Jarrell, J. A., Gupta, R., Balboni, I., Chakravarty, E. F., Sokolove, J., Shum, A. K., Anderson, M. S., Cheng, M. H., Robinson, W. H., Browne, S. K., Holland, S. M., Baechler, E. C., Utz, P. J. 2013; 123 (12): 5135-5145

  Abstract

  Autoantibodies against cytokines, chemokines, and growth factors inhibit normal immunity and are implicated in inflammatory autoimmune disease and diseases of immune deficiency. In an effort to evaluate serum from autoimmune and immunodeficient patients for Abs against cytokines, chemokines, and growth factors in a high-throughput and unbiased manner, we constructed a multiplex protein microarray for detection of serum factor-binding Abs and used the microarray to detect autoantibody targets in SLE. We designed a nitrocellulose-surface microarray containing human cytokines, chemokines, and other circulating proteins and demonstrated that the array permitted specific detection of serum factor-binding probes. We used the arrays to detect previously described autoantibodies against cytokines in samples from individuals with autoimmune polyendocrine syndrome type 1 and chronic mycobacterial infection. Serum profiling from individuals with SLE revealed that among several targets, elevated IgG autoantibody reactivity to B cell-activating factor (BAFF) was associated with SLE compared with control samples. BAFF reactivity correlated with the severity of disease-associated features, including IFN-α-driven SLE pathology. Our results showed that serum factor protein microarrays facilitate detection of autoantibody reactivity to serum factors in human samples and that BAFF-reactive autoantibodies may be associated with an elevated inflammatory disease state within the spectrum of SLE.

  View details for DOI 10.1172/JCI70231

  View details for Web of Science ID 000327826100020

  View details for PubMedID 24270423

  View details for PubMedCentralID PMC3859403

• Serum autoantibodies to myelin peptides distinguish acute disseminated encephalomyelitis from relapsing- remitting multiple sclerosis. Multiple sclerosis (Houndmills, Basingstoke, England) Van Haren, K., Tomooka, B. H., Kidd, B. A., Banwell, B., Bar-Or, A., Chitnis, T., Tenembaum, S. N., Pohl, D., Rostasy, K., Dale, R. C., O'Connor, K. C., Hafler, D. A., Steinman, L., Robinson, W. H. 2013; 19 (13): 1726-1733

  Abstract

  BACKGROUND AND OBJECTIVE: Acute disseminated encephalomyelitis (ADEM) and relapsing-remitting multiple sclerosis (RRMS) share overlapping clinical, radiologic and laboratory features at onset. Because autoantibodies may contribute to the pathogenesis of both diseases, we sought to identify autoantibody biomarkers that are capable of distinguishing them. METHODS: We used custom antigen arrays to profile anti-myelin-peptide autoantibodies in sera derived from individuals with pediatric ADEM (n = 15), pediatric multiple sclerosis (Ped MS; n = 11) and adult MS (n = 15). Using isotype-specific secondary antibodies, we profiled both IgG and IgM reactivities. We used Statistical Analysis of Microarrays software to confirm the differences in autoantibody reactivity profiles between ADEM and MS samples. We used Prediction Analysis of Microarrays software to generate and validate prediction algorithms, based on the autoantibody reactivity profiles. RESULTS: ADEM was characterized by IgG autoantibodies targeting epitopes derived from myelin basic protein, proteolipid protein, myelin-associated oligodendrocyte basic glycoprotein, and alpha-B-crystallin. In contrast, MS was characterized by IgM autoantibodies targeting myelin basic protein, proteolipid protein, myelin-associated oligodendrocyte basic glycoprotein and oligodendrocyte-specific protein. We generated and validated prediction algorithms that distinguish ADEM serum (sensitivity 62-86%; specificity 56-79%) from MS serum (sensitivity 40-87%; specificity 62-86%) on the basis of combined IgG and IgM anti-myelin autoantibody reactivity to a small number of myelin peptides. CONCLUSIONS: Combined profiles of serum IgG and IgM autoantibodies identified myelin antigens that may be useful for distinguishing MS from ADEM. Further studies are required to establish clinical utility. Further biological assays are required to delineate the pathogenic potential of these antibodies.

  View details for DOI 10.1177/1352458513485653

  View details for PubMedID 23612879

• Autoimmune Attack Takes Your Breath Away SCIENCE TRANSLATIONAL MEDICINE Lindstrom, T. M., Robinson, W. H. 2013; 5 (206): 206fs36

  Abstract

  Autoimmune targeting of a lung-specific protein can cause interstitial lung disease (Shum et al., this issue).

  View details for PubMedID 24107774

• Novel Biomarkers From Peripheral Blood Mononuclear Cells Indicate Disease Activity In Rheumatoid Arthritis Patients. Ptacek, J., Hawtin, R., Louie, B., Evensen, E., Cordeiro, J., Mittleman, B., Atallah, M., Cesano, A., Bingham, C. O., Cofield, S., Curtis, J. R., Danila, M. I., Furie, R. A., Genovese, M. C., Levesque, M. C., Moreland, L. W., Nigrovic, P. A., O'Dell, J. R., Robinson, W. H., Shadick, N. A., St Clair, E., Striebich, C. C., Thiele, G. M., Gregersen, P. K., Bridges, S. WILEY-BLACKWELL. 2013: S974–S975

  View details for Web of Science ID 000325359205235

• IgM Rheumatoid Factor As a Potentiator Of Anti-Citrullinated Protein Antibody Mediated Inflammation In Rheumatoid Arthritis Sokolove, J., Cheng, D., Johnson, D. S., Mikuls, T. R., Robinson, W. H. WILEY-BLACKWELL. 2013: S343–S344

  View details for Web of Science ID 000325359202314

• Alveolar Bone Loss Is Associated With Disease Activity and ACPA Expression In Rheumatoid Arthritis Mikuls, T. R., Payne, J., Yu, F., Thiele, G. M., Gonzalez, S., Markt, J., Sokolove, J., Robinson, W. H., Reynolds, R. J., Cannon, G. W., McGowan, D., Kerr, G. S., Redman, R., Reimold, A. M., Griffiths, G., Beatty, M., Schmid, M., Johnson, P., Bergman, D., Hamilton, B. C., Erickson, A. R., O'Dell, J. R. WILEY-BLACKWELL. 2013: S345

  View details for Web of Science ID 000325359202317

• Role Of B Cells and/Or Autoantibodies In Pulmonary Manifestations Of Inflammatory Arthritis Peterson, L. K., Sokolove, J., Jedlicka, P., Lahey, L. J., Robinson, W. H., Dragone, L. L. WILEY-BLACKWELL. 2013: S395–S396

  View details for Web of Science ID 000325359202427

• Decreased Plasma Levels Of Soluble CD18 Link Leukocyte Migration and Disease Activity In Spondyloarthritis Kragstrup, T. W., Jalilian, B., Hvid, M., Ostgard, R., Schiottz-Christensen, B., Jurik, A. G., Robinson, W. H., Vorup-Jensen, T., Deleuran, B. WILEY-BLACKWELL. 2013: S401

  View details for Web of Science ID 000325359202441

• Chemokine Receptor Polymorphisms On Chromosome 3 Are Associated With Anticitrullinated Protein Antibody Specificities In African Americans With Rheumatoid Arthritis Reynolds, R. J., Danila, M. I., Sokolove, J., Robinson, W. H., Conn, D. L., Jonas, B. L., Callahan, L. F., Moreland, L. W., Brasington, R. D., Smith, E. A., Gregersen, P. K., Bridges, S. WILEY-BLACKWELL. 2013: S805

  View details for Web of Science ID 000325359204349

• Tumor Necrosis Factor Alpha Induces Anti-Citrullinated Protein Antibodies and Arthritis In Part Through Peptidyl Arginine Deiminase 4 Shelef, M. A., Sokolove, J., Wagner, C., Sackmann, E., Warner, T., Beebe, D., Robinson, W. H., Huttenlocher, A. WILEY-BLACKWELL. 2013: S947

  View details for Web of Science ID 000325359205171

• Recombinant Monoclonal Antibodies Derived From Single CD19+Synovial B Cells Of RA Patients With Tertiary Lymphoid Structures Display a Strong Immunoreactivity Towards Citrullinated Histones Corsiero, E., Bombardieri, M., Carlotti, E., Wardemann, H., Robinson, W. H., Pitzalis, C. WILEY-BLACKWELL. 2013: S1127–S1128

  View details for Web of Science ID 000325359206077

• Antibodies To Citrullinated Enolase, Fibrinogen, and Vimentin Are Associated With Markers Of Endothelial Dysfunction In First-Degree Relatives Of Patients With Rheumatoid Arthritis: The Studies Of The Etiology Of Rheumatoid Arthritis Hughes-Austin, J. M., Young, K. A., Deane, K. D., Weisman, M. H., Buckner, J. H., Mikuls, T. R., O'Dell, J. R., Keating, R. M., Gregersen, P. K., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M. WILEY-BLACKWELL. 2013: S1133–S1134

  View details for Web of Science ID 000325359206089

• Carboxypeptidase B2 Down-Regulates Osteoclast Activation In Inflammation. Song, J., Lee, S., Park, Y., Lee, S., Leung, L., Robinson, W. H. WILEY-BLACKWELL. 2013: S781

  View details for Web of Science ID 000325359204288

• The Relationship Between Anti-Citrullinated Protein Antibodies and Radiographic Damage In African-Americans With Rheumatoid Arthritis Danila, M. I., Reynolds, R. J., Aslibekyan, S., Tamhane, A., Hughes, L. B., Sokolove, J., Robinson, W. H., Conn, D. L., Jonas, B. L., Callahan, L. F., Moreland, L. W., Brasington, R. D., Smith, E. A., Mikuls, T. R., van der Heijde, D., Bridges, S. WILEY-BLACKWELL. 2013: S1028

  View details for Web of Science ID 000325359205352

• Prediction Of TNF Inhibitor Response In Rheumatoid Arthritis Patients Using Single Cell Network Profiling Of Intracellular Immune Signaling Ptacek, J., Hawtin, R., Louie, B., Evensen, E., Cordeiro, J., Mittleman, B., Atallah, M., Cesano, A., Bingham, C. O., Cofield, S., Curtis, J. R., Danila, M. I., Furie, R. A., Genovese, M. C., Levesque, M. C., Moreland, L. W., Nigrovic, P. A., O'Dell, J. R., Robinson, W. H., Shadick, N. A., St Clair, E., Striebich, C. C., Thiele, G. M., Gregersen, P. K., Bridges, S. WILEY-BLACKWELL. 2013: S375

  View details for Web of Science ID 000325359202381

• Whole-genome sequencing identifies a recurrent functional synonymous mutation in melanoma PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Gartner, J. J., Parker, S. C., Prickett, T. D., Dutton-Regester, K., Stitzel, M. L., Lin, J. C., Davis, S., Simhadri, V. L., Jha, S., Katagiri, N., Gotea, V., Teer, J. K., Wei, X., Morken, M. A., Bhanot, U. K., Chen, G., Elnitski, L. L., Davies, M. A., Gershenwald, J. E., Carter, H., Karchin, R., Robinson, W., Robinson, S., Rosenberg, S. A., Collins, F. S., Parmigiani, G., Komar, A. A., Kimchi-Sarfaty, C., Hayward, N. K., Margulies, E. H., Samuels, Y. 2013; 110 (33): 13481-13486

  Abstract

  Synonymous mutations, which do not alter the protein sequence, have been shown to affect protein function [Sauna ZE, Kimchi-Sarfaty C (2011) Nat Rev Genet 12(10):683-691]. However, synonymous mutations are rarely investigated in the cancer genomics field. We used whole-genome and -exome sequencing to identify somatic mutations in 29 melanoma samples. Validation of one synonymous somatic mutation in BCL2L12 in 285 samples identified 12 cases that harbored the recurrent F17F mutation. This mutation led to increased BCL2L12 mRNA and protein levels because of differential targeting of WT and mutant BCL2L12 by hsa-miR-671-5p. Protein made from mutant BCL2L12 transcript bound p53, inhibited UV-induced apoptosis more efficiently than WT BCL2L12, and reduced endogenous p53 target gene transcription. This report shows selection of a recurrent somatic synonymous mutation in cancer. Our data indicate that silent alterations have a role to play in human cancer, emphasizing the importance of their investigation in future cancer genome studies.

  View details for DOI 10.1073/pnas.1304227110

  View details for Web of Science ID 000323069200063

  View details for PubMedID 23901115

  View details for PubMedCentralID PMC3746936

• Relatives Without Rheumatoid Arthritis Show Reactivity to Anti-Citrullinated Protein/Peptide Antibodies That Are Associated With Arthritis-Related Traits: Studies of the Etiology of Rheumatoid Arthritis ARTHRITIS AND RHEUMATISM Young, K. A., Deane, K. D., Derber, L. A., Hughes-Austin, J. M., Wagner, C. A., Sokolove, J., Weisman, M. H., Buckner, J. H., Mikuls, T. R., O'Dell, J. R., Keating, R. M., Gregersen, P. K., Robinson, W. H., Holers, V. M., Norris, J. M. 2013; 65 (8): 1995-2004

  Abstract

  To examine reactivity to anti-citrullinated protein/peptide antibodies (ACPAs) and determine associations between ACPAs and other rheumatoid arthritis (RA)-related autoantibodies and clinically assessed swollen or tender joints in unaffected first-degree relatives of RA patients.Serum samples were obtained from first-degree relatives without RA according to the 1987 American College of Rheumatology (ACR) and the 2010 ACR/European League Against Rheumatism classification criteria. A bead-based assay was used to measure 16 separate ACPAs in sera from 111 antibody-positive first-degree relatives who were positive on at least 1 visit for any of 5 RA-related autoantibodies (rheumatoid factor [RF], anti-cyclic citrullinated peptide 2 [anti-CCP-2], and RF isotypes), and sera from 99 antibody-negative first-degree relatives who were never autoantibody positive. Cutoffs for positivity for each ACPA were determined using receiver operating characteristic curves derived from data on 200 RA patients and 98 blood donor controls, in which positivity for ≥9 ACPAs had 92% specificity and 62% sensitivity for RA. In first-degree relatives, ACPA reactivity was assessed, and associations between ACPAs (number positive, and positivity for ≥9 ACPAs) and RA-related characteristics were examined.Fifty-seven percent of anti-CCP-2-positive first-degree relatives and 8% of anti-CCP-2- negative first-degree relatives were positive for ≥9 ACPAs. After adjusting for age, sex, ethnicity, and pack-years of smoking, an increasing number of ACPAs was directly associated with the presence of ≥1 tender joint on examination (odds ratio [OR] 1.18, 95% confidence interval [95% CI] 1.04-1.34), with the greatest risk of having ≥1 tender joint seen in first-degree relatives positive for ≥9 ACPAs (OR 5.00, 95% CI 1.37-18.18).RA-free first-degree relatives (even those negative for RF and anti-CCP-2) demonstrate reactivity to multiple ACPAs, and the presence of an increasing number of ACPAs may be associated with signs of joint inflammation. Prospective evaluation of the relationship between these findings and the progression of classifiable RA is warranted.

  View details for DOI 10.1002/art.38022

  View details for Web of Science ID 000322329100008

  View details for PubMedID 23754702

  View details for PubMedCentralID PMC3729718

• Brief report: citrullination within the atherosclerotic plaque: a potential target for the anti-citrullinated protein antibody response in rheumatoid arthritis. Arthritis and rheumatism Sokolove, J., Brennan, M. J., Sharpe, O., Lahey, L. J., Kao, A. H., Krishnan, E., Edmundowicz, D., Lepus, C. M., Wasko, M. C., Robinson, W. H. 2013; 65 (7): 1719-1724

  Abstract

  BACKGROUND/PURPOSE: Patients with rheumatoid arthritis (RA) are at increased risk for cardiovascular disease, an observation not explained by traditional cardiac risk factors and generally limited to those with RA-associated autoantibodies such as rheumatoid factor and anti-citrullinated protein antibodies (ACPA). We hypothesized that citrullinated proteins within the atherosclerotic plaque can be targeted by ACPA, forming stimulatory immune complexes which propagate the progression of atherosclerosis. METHODS AND RESULTS: Protein lysates prepared from atherosclerotic segments of human aorta were investigated for the presence of citrulline-modified proteins and specifically citrullinated fibrinogen (cFb) by immunoprecipitation and/or immunoblotting followed by mass spectrometry. Immunohistochemistry was performed in coronary artery plaques for the presence of citrullinated proteins and the PAD4 enzyme. Serum levels of anti-cyclic citrullinated peptide (CCP), anti-citrullinated vimentin (cVim), and anti-cFb antibodies were measured in 134 women with seropositive RA previously characterized for the presence of subclinical atherosclerosis by electron beam CT scan (EBCT). Western analysis of atherosclerotic plaque lysates demonstrated several citrullinated proteins and the presence of cFb was confirmed by immunoprecipitation, and mass spectrometry. Immunohistochemistry demonstrated co-localization of citrullinated proteins and the PAD4 enzyme within the coronary artery plaque. In age-adjusted regression models, antibodies targeting cFb and cit-vimentin, but not CCP2, were associated with an increased aortic plaque burden. CONCLUSION: Citrullinated proteins are prevalent within the atherosclerotic plaque, and certain ACPAs are associated with atherosclerotic burden. These observations suggest that targeting of citrullinated epitopes, specifically cFb, within the atherosclerotic plaque could provide a mechanism for accelerated atherosclerosis observed in patients with RA. © 2013 American College of Rheumatology.

  View details for DOI 10.1002/art.37961

  View details for PubMedID 23553485

• Citrullination Within the Atherosclerotic Plaque: A Potential Target for the Anti-Citrullinated Protein Antibody Response in Rheumatoid Arthritis ARTHRITIS AND RHEUMATISM Sokolove, J., Brennan, M. J., Sharpe, O., Lahey, L. J., Kao, A. H., Krishnan, E., Edmundowicz, D., Lepus, C. M., Wasko, M. C., Robinson, W. H. 2013; 65 (7): 1719-1724

  Abstract

  BACKGROUND/PURPOSE: Patients with rheumatoid arthritis (RA) are at increased risk for cardiovascular disease, an observation not explained by traditional cardiac risk factors and generally limited to those with RA-associated autoantibodies such as rheumatoid factor and anti-citrullinated protein antibodies (ACPA). We hypothesized that citrullinated proteins within the atherosclerotic plaque can be targeted by ACPA, forming stimulatory immune complexes which propagate the progression of atherosclerosis. METHODS AND RESULTS: Protein lysates prepared from atherosclerotic segments of human aorta were investigated for the presence of citrulline-modified proteins and specifically citrullinated fibrinogen (cFb) by immunoprecipitation and/or immunoblotting followed by mass spectrometry. Immunohistochemistry was performed in coronary artery plaques for the presence of citrullinated proteins and the PAD4 enzyme. Serum levels of anti-cyclic citrullinated peptide (CCP), anti-citrullinated vimentin (cVim), and anti-cFb antibodies were measured in 134 women with seropositive RA previously characterized for the presence of subclinical atherosclerosis by electron beam CT scan (EBCT). Western analysis of atherosclerotic plaque lysates demonstrated several citrullinated proteins and the presence of cFb was confirmed by immunoprecipitation, and mass spectrometry. Immunohistochemistry demonstrated co-localization of citrullinated proteins and the PAD4 enzyme within the coronary artery plaque. In age-adjusted regression models, antibodies targeting cFb and cit-vimentin, but not CCP2, were associated with an increased aortic plaque burden. CONCLUSION: Citrullinated proteins are prevalent within the atherosclerotic plaque, and certain ACPAs are associated with atherosclerotic burden. These observations suggest that targeting of citrullinated epitopes, specifically cFb, within the atherosclerotic plaque could provide a mechanism for accelerated atherosclerosis observed in patients with RA. © 2013 American College of Rheumatology.

  View details for DOI 10.1002/art.37961

  View details for Web of Science ID 000322155000007

• Plasmid-Encoded Proinsulin Preserves C-Peptide While Specifically Reducing Proinsulin-Specific CD8+ T Cells in Type 1 Diabetes. Science translational medicine Roep, B. O., Solvason, N., Gottlieb, P. A., Abreu, J. R., Harrison, L. C., Eisenbarth, G. S., Yu, L., Leviten, M., Hagopian, W. A., Buse, J. B., von Herrath, M., Quan, J., King, R. S., Robinson, W. H., Utz, P. J., Garren, H., Steinman, L. 2013; 5 (191): 191ra82-?

  Abstract

  In type 1 diabetes (T1D), there is an intense inflammatory response that destroys the β cells in the pancreatic islets of Langerhans, the site where insulin is produced and released. A therapy for T1D that targets the specific autoimmune response in this disease while leaving the remainder of the immune system intact, has long been sought. Proinsulin is a major target of the adaptive immune response in T1D. We hypothesized that an engineered DNA plasmid encoding proinsulin (BHT-3021) would preserve β cell function in T1D patients through reduction of insulin-specific CD8(+) T cells. We studied 80 subjects over 18 years of age who were diagnosed with T1D within the past 5 years. Subjects were randomized 2:1 to receive intramuscular injections of BHT-3021 or BHT-placebo, weekly for 12 weeks, and then monitored for safety and immune responses in a blinded fashion. Four dose levels of BHT-3021 were evaluated: 0.3, 1.0, 3.0, and 6.0 mg. C-peptide was used both as an exploratory efficacy measure and as a safety measure. Islet-specific CD8(+) T cell frequencies were assessed with multimers of monomeric human leukocyte antigen class I molecules loaded with peptides from pancreatic and unrelated antigens. No serious adverse events related to BHT-3021 were observed. C-peptide levels improved relative to placebo at all doses, at 1 mg at the 15-week time point (+19.5% BHT-3021 versus -8.8% BHT-placebo, P < 0.026). Proinsulin-reactive CD8(+) T cells, but not T cells against unrelated islet or foreign molecules, declined in the BHT-3021 arm (P < 0.006). No significant changes were noted in interferon-γ, interleukin-4 (IL-4), or IL-10 production in CD4 T cells. Thus, we demonstrate that a plasmid encoding proinsulin reduces the frequency of CD8(+) T cells reactive to proinsulin while preserving C-peptide over the course of dosing.

  View details for DOI 10.1126/scitranslmed.3006103

  View details for PubMedID 23803704

• Multiple cytokines and chemokines are associated with rheumatoid arthritis-related autoimmunity in first-degree relatives without rheumatoid arthritis: Studies of the Aetiology of Rheumatoid Arthritis (SERA) ANNALS OF THE RHEUMATIC DISEASES Hughes-Austin, J. M., Deane, K. D., Derber, L. A., Kolfenbach, J. R., Zerbe, G. O., Sokolove, J., Lahey, L. J., Weisman, M. H., Buckner, J. H., Mikuls, T. R., O'Dell, J. R., Keating, R. M., Gregersen, P. K., Robinson, W. H., Holers, V. M., Norris, J. M. 2013; 72 (6): 901-907

  Abstract

  We investigated whether rheumatoid arthritis (RA)-related autoantibodies were associated with systemic inflammation in a prospective cohort of first-degree relatives (FDRs) of RA probands, a population without RA but at increased risk for its future development.We studied 44 autoantibody positive FDRs, of whom 29 were rheumatoid factor (RF) positive, 25 were positive for the high risk autoantibody profile (HRP), that is, positive for anti-cyclic citrullinated peptide and/or for at least two RF IgM, IgG or IgA isotypes, and nine FDRs who were positive for both; and 62 FDRs who were never autoantibody positive. Twenty-five cytokines/chemokines were measured using a bead-based assay in serum. As a comprehensive measure of inflammation, we calculated a Cytokine Score by summing all cytokine/chemokine levels, weighted by their regression coefficients for RA-autoantibody association. We compared C-reactive protein, individual cytokines/chemokines and Cytokine Score to the outcomes: positivity for RF and for the HRP using logistic regression.Adjusting for age, sex, ethnicity and ever smoking, the Cytokine Score and levels of IL-6 and IL-9 were associated with both RF and HRP. IL-2, granulocyte macrophage-colony stimulating factor (GM-CSF), and interferon (IFN)-γ were associated with HRP only. Associations between the Cytokine Score and RF and HRP positivity were replicated in an independent military personnel cohort.In first-degree relatives of patients with RA, RA-related autoimmunity is associated with inflammation, as evidenced by associations with multiple cytokines and chemokines.

  View details for DOI 10.1136/annrheumdis-2012-201505

  View details for Web of Science ID 000318907100024

  View details for PubMedID 22915618

  View details for PubMedCentralID PMC3726193

• Predictive Value of Autoantibody Testing for Validating Self-reported Diagnoses of Rheumatoid Arthritis in the Women's Health Initiative AMERICAN JOURNAL OF EPIDEMIOLOGY Walitt, B., Mackey, R., Kuller, L., Deane, K. D., Robinson, W., Holers, V. M., Chang, Y., Moreland, L. 2013; 177 (9): 887-893

  Abstract

  Rheumatoid arthritis (RA) research using large databases is limited by insufficient case validity. Of 161,808 postmenopausal women in the Women's Health Initiative, 15,691 (10.2%) reported having RA, far higher than the expected 1% population prevalence. Since chart review for confirmation of an RA diagnosis is impractical in large cohort studies, the current study (2009-2011) tested the ability of baseline serum measurements of rheumatoid factor and anti-cyclic citrullinated peptide antibodies, second-generation assay (anti-CCP2), to identify physician-validated RA among the chart-review study participants with self-reported RA (n = 286). Anti-CCP2 positivity had the highest positive predictive value (PPV) (80.0%), and rheumatoid factor positivity the lowest (44.6%). Together, use of disease-modifying antirheumatic drugs and anti-CCP2 positivity increased PPV to 100% but excluded all seronegative cases (approximately 15% of all RA cases). Case definitions inclusive of seronegative cases had PPVs between 59.6% and 63.6%. False-negative results were minimized in these test definitions, as evidenced by negative predictive values of approximately 90%. Serological measurements, particularly measurement of anti-CCP2, improved the test characteristics of RA case definitions in the Women's Health Initiative.

  View details for DOI 10.1093/aje/kws310

  View details for Web of Science ID 000318576300006

  View details for PubMedID 23492764

  View details for PubMedCentralID PMC4023293

• Mechanistic biomarkers for clinical decision making in rheumatic diseases NATURE REVIEWS RHEUMATOLOGY Robinson, W. H., Lindstrom, T. M., Cheung, R. K., Sokolove, J. 2013; 9 (5): 267-276

  Abstract

  The use of biomarkers is becoming increasingly intrinsic to the practice of medicine and holds great promise for transforming the practice of rheumatology. Biomarkers have the potential to aid clinical diagnosis when symptoms are present or to provide a means of detecting early signs of disease when they are not. Some biomarkers can serve as early surrogates of eventual clinical outcomes or guide therapeutic decision making by enabling identification of individuals likely to respond to a specific therapy. Using biomarkers might reduce the costs of drug development by enabling individuals most likely to respond to be enrolled in clinical trials, thereby minimizing the number of participants required. In this Review, we discuss the current use and the potential of biomarkers in rheumatology and in select fields at the forefront of biomarker research. We emphasize the value of different types of biomarkers, addressing the concept of 'actionable' biomarkers, which can be used to guide clinical decision making, and 'mechanistic' biomarkers, a subtype of actionable biomarker that is embedded in disease pathogenesis and, therefore, represents a potentially superior biomarker. We provide examples of actionable and mechanistic biomarkers currently available, and discuss how development of such biomarkers could revolutionize clinical practice and drug development.

  View details for DOI 10.1038/nrrheum.2013.14

  View details for Web of Science ID 000318482100004

  View details for PubMedID 23419428

  View details for PubMedCentralID PMC3673766

• Optimal approaches to data collection and analysis of potential immune mediated disorders in clinical trials of new vaccines VACCINE Da Silva, F. T., De Keyser, F., Lambert, P., Robinson, W. H., Westhovens, R., Sindic, C. 2013; 31 (14): 1870-1876

  Abstract

  The potential for development of autoimmune diseases after vaccination with new vaccines containing novel adjuvants is a theoretical concern. Randomised, placebo-controlled trials are the best method for assessing a potential causal relationship between an adverse event and vaccination, but usually have a sample size too small to detect adverse events occurring in <1% of subjects. Incomplete case documentation may hamper definitive diagnoses, preventing accurate causality assessment. To date there are no guidelines for collection, documentation and monitoring of potential immune mediated disorders (pIMD) reported in the course of clinical trials with adjuvanted vaccines.This paper proposes a methodology for collection of pIMDs in clinical vaccine trials, with the objective of obtaining complete and reliable data using standardised methodology for its collection and analysis.The role of the study investigator in prospective, standardised safety data collection is key and can be facilitated by providing a pIMD list in study documents and disease-specific standard questionnaires to assist timely and thorough documentation. External expert review of histopathology samples or other specialised diagnostic data would increase diagnostic accuracy. Centralised case ascertainment using standard case definitions would identify true cases of interest. We propose collection of safety data for at least 6 months and up to one year after the last vaccine dose. Bio-banking as a platform for collecting samples from enrolled patients for future use (e.g., to measure biomarkers of diagnostic, prognostic or predictive utility) could eventually provide valuable information in cases where a pIMD is diagnosed during the study period.Standardised collection of safety data to allow appropriate analyses are optimal approaches for detecting rare events in clinical trials. Appropriate data analysis will then more reliably define potential causal relationships with vaccination.

  View details for DOI 10.1016/j.vaccine.2013.01.042

  View details for Web of Science ID 000317450200016

  View details for PubMedID 23391600

• The exposure of autoantigens by microparticles underlies the formation of potent inflammatory components: the microparticle-associated immune complexes EMBO MOLECULAR MEDICINE Cloutier, N., Tan, S., Boudreau, L. H., Cramb, C., Subbaiah, R., Lahey, L., Albert, A., Shnayder, R., Gobezie, R., Nigrovic, P. A., Farndale, R. W., Robinson, W. H., Brisson, A., Lee, D. M., Boilard, E. 2013; 5 (2): 235-249

  Abstract

  Immunoglobulins, antigens and complement can assemble to form immune complexes (IC). ICs can be detrimental as they propagate inflammation in autoimmune diseases. Like ICs, submicron extracellular vesicles termed microparticles (MP) are present in the synovial fluid from patients affected with autoimmune arthritis. We examined MPs in rheumatoid arthritis (RA) using high sensitivity flow cytometry and electron microscopy. We find that the MPs in RA synovial fluid are highly heterogeneous in size. The observed larger MPs were in fact MP-containing ICs (mpICs) and account for the majority of the detectable ICs. These mpICs frequently express the integrin CD41, consistent with platelet origin. Despite expression of the Fc receptor FcγRIIa by platelet-derived MPs, we find that the mpICs form independently of this receptor. Rather, mpICs display autoantigens vimentin and fibrinogen, and recognition of these targets by anti-citrullinated peptide antibodies contributes to the production of mpICs. Functionally, platelet mpICs are highly pro-inflammatory, eliciting leukotriene production by neutrophils. Taken together, our data suggest a unique role for platelet MPs as autoantigen-expressing elements capable of perpetuating formation of inflammatory ICs.

  View details for DOI 10.1002/emmm.201201846

  View details for Web of Science ID 000314661800010

  View details for PubMedID 23165896

  View details for PubMedCentralID PMC3569640

• Anti-citrullinated peptide autoantibodies, human leukocyte antigen shared epitope and risk of future rheumatoid arthritis: a nested case-control study ARTHRITIS RESEARCH & THERAPY Arkema, E. V., Goldstein, B. L., Robinson, W., Sokolove, J., Wagner, C. A., Malspeis, S., Rosner, B., Grodstein, F., Karlson, E. W., Costenbader, K. H. 2013; 15 (5)

  Abstract

  The aim of this study was to characterize anti-citrullinated peptide antibody (ACPA) serostatus in pre-clinical rheumatoid arthritis (RA) with and without Human Leukocyte Antigen-Shared Epitope (HLA-SE) alleles.We identified 192 women in the Nurses' Health Study cohorts with blood samples obtained 4 months to 17 years prior to medical record-confirmed RA diagnosis. Three controls were selected matched on age, cohort, menopausal status and post-menopausal hormone use. Reactivities to 18 ACPAs were measured using a custom BioPlex platform. We used conditional logistic regression to calculate the relative risk (RR) of RA for any ACPA-positive and peptide-specific ACPA-positive and examined RRs by time between blood draw and RA onset. Measures of multiplicative and additive interaction between any ACPA-positive and HLA-SE were calculated.All ACPAs by peptide groups were significantly associated with RA risk, RRs ranged from 4.7 to 11.7. The association between ACPA and RA varied over time with the strongest association in those with blood draw less than 5 years before onset (RR 17.0 [95% CI 5.8 to 53.7]) and no association 10 or more years prior to onset (RR 1.4 [95% CI 0.5 to 4.3]). Individuals with both HLA-SE and any ACPA-positive had the highest risk of RA. HLA-SE-positive RA cases showed reactivity to more ACPA types than HLA-SE negative (χ2 test for trend, P = 0.01).There is increasing ACPA reactivity up to 10 years before RA onset with the strongest association within 5 years of RA onset. The magnitude of the response to ACPAs, in combination with the presence of HLA-SE, is most important for identifying those individuals with the highest risk of RA.

  View details for DOI 10.1186/ar4342

  View details for Web of Science ID 000329737400059

  View details for PubMedID 24286474

  View details for PubMedCentralID PMC3953952

• Activation of Classical Complement Pathway is Associated with the Development of Idiopathic Pulmonary Arterial Hypertension (IPAH) Saito, T., Sharpe, O., Harada, D., Tamosiuniene, R., Li, C. G., Nicolls, M., Robinson, W. H., Rabinovitch, M. LIPPINCOTT WILLIAMS & WILKINS. 2012

  View details for Web of Science ID 000208885006195

• Porphyromonas gingivalis and Disease-Related Autoantibodies in Individuals at Increased Risk of Rheumatoid Arthritis ARTHRITIS AND RHEUMATISM Mikuls, T. R., Thiele, G. M., Deane, K. D., Payne, J. B., O'Dell, J. R., Yu, F., Sayles, H., Weisman, M. H., Gregersen, P. K., Buckner, J. H., Keating, R. M., Derber, L. A., Robinson, W. H., Holers, V. M., Norris, J. M. 2012; 64 (11): 3522-3530

  Abstract

  To examine the relationship of Porphyromonas gingivalis to the presence of autoantibodies in individuals at risk of rheumatoid arthritis (RA).Study participants included the following: 1) a cohort enriched in subjects with HLA-DR4 and 2) subjects at risk of RA by virtue of having a first-degree relative with RA. None of the study subjects satisfied the American College of Rheumatology 1987 classification criteria for RA. Autoantibodies measured included anti-citrullinated protein antibody (ACPA; by second-generation anti-cyclic citrullinated peptide antibody enzyme-linked immunosorbent assay [ELISA]) and rheumatoid factor (RF; by nephelometry or ELISA for IgA, IgM, or IgG isotype). Individuals were considered autoantibody positive (n = 113) if they had ≥1 RA-related autoantibody; individuals were further categorized as high risk (n = 38) if they had ACPA or positive findings ≥2 assays for RF. Autoantibody-negative individuals (n = 171) served as a comparator group. Antibody to P gingivalis, P intermedia, and F nucleatum were measured. Associations of bacterial antibodies with group status were examined using logistic regression.Anti-P gingivalis concentrations were higher in high-risk (P = 0.011) and autoantibody positive group (P = 0.010) than in the autoantibody negative group. There were no group differences in anti-P intermedia or anti-F nucleatum concentrations. After multivariable adjustment, anti-P gingivalis concentrations (but not anti-P intermedia or anti-F nucleatum) were significantly associated with autoantibody-positive and high-risk status (P < 0.05).Immunity to P gingivalis, but not P intermedia or F nucleatum, is significantly associated with the presence of RA-related autoantibodies in individuals at risk of RA. These results support the hypothesis that infection with P gingivalis may play a central role in the early loss of tolerance to self antigens that occurs in the pathogenesis of RA.

  View details for DOI 10.1002/art.34595

  View details for Web of Science ID 000310544500006

  View details for PubMedID 22736291

  View details for PubMedCentralID PMC3467347

• Oxidative stress-responsive microRNA-320 regulates glycolysis in diverse biological systems FASEB JOURNAL Tang, H., Lee, M., Sharpe, O., Salamone, L., Noonan, E. J., Hoang, C. D., Levine, S., Robinson, W. H., Shrager, J. B. 2012; 26 (11): 4710-4721

  Abstract

  Glycolysis is the initial step of glucose catabolism and is up-regulated in cancer cells (the Warburg Effect). Such shifts toward a glycolytic phenotype have not been explored widely in other biological systems, and the molecular mechanisms underlying the shifts remain unknown. With proteomics, we observed increased glycolysis in disused human diaphragm muscle. In disused muscle, lung cancer, and H(2)O(2)-treated myotubes, we show up-regulation of the rate-limiting glycolytic enzyme muscle-type phosphofructokinase (PFKm, >2 fold, P<0.05) and accumulation of lactate (>150%, P<0.05). Using microRNA profiling, we identify miR-320a as a regulator of PFKm expression. Reduced miR-320a levels (to ∼50% of control, P<0.05) are associated with the increased PFKm in each of these diverse systems. Manipulation of miR-320a levels both in vitro and in vivo alters PFKm and lactate levels in the expected directions. Further, miR-320a appears to regulate oxidative stress-induced PFKm expression, and reduced miR-320a allows greater induction of glycolysis in response to H(2)O(2) treatment. We show that this microRNA-mediated regulation occurs through PFKm's 3' untranslated region and that Ets proteins are involved in the regulation of PFKm via miR-320a. These findings suggest that oxidative stress-responsive microRNA-320a may regulate glycolysis broadly within nature.

  View details for DOI 10.1096/fj.11-197467

  View details for PubMedID 22767230

• The Associations of HLA-DR Shared Epitope Alleles and Serum Cytokines Among Postmenopausal Women with Rheumatoid Factor and Anti-Citrullinated Protein Antibody-Positive Rheumatoid Arthritis. Annual Scientific Meeting of the American-College-of-Rheumatology (ACR) and Association-of-Rheumatology-Health-Professionals (ARHP) Talabi, M. B., Mackey, R., Moreland, L. W., Dorman, J., Deane, K. D., Sokolove, J., Holers, V. M., Robinson, W. H., Wallitt, B., Kuller, L. WILEY-BLACKWELL. 2012: S895–S895

  View details for Web of Science ID 000309748305036

• Differential Expression of Hepatocyte Growth Factor (HGF) in Patients with Systemic Sclerosis-Associated Pulmonary Arterial Hypertension Chung, L., Cramb, C., Robinson, W. H., Steen, V. D., Zamanian, R. T. WILEY-BLACKWELL. 2012: S632

  View details for Web of Science ID 000309748303192

• Smoking Status Is Associated with Inflammatory Cytokine Profile and Disease Activity in Anti-Citrullinated Protein Antibody Positive Rheumatoid Arthritis: Decreased Inflammation and Disease Improvement with Smoking Cessation? Cramb, C., Sokolove, J., Thiele, G. M., Kerr, G. S., Cannon, G. W., Reimold, A. M., Mikuls, T. R., Robinson, W. H. WILEY-BLACKWELL. 2012: S508–S509

  View details for Web of Science ID 000309748302262

• Citrullination within the Atherosclerotic Plaque: A New Potential Target for Anti-Citrullinated Protein Antibodies Sokolove, J., Share, O., Brennan, M., Lahey, L. J., Kao, A. H., Krishnan, E., Wasko, M., Robinson, W. H. WILEY-BLACKWELL. 2012: S723

  View details for Web of Science ID 000309748303411

• Adiponectin Is Associated with Pro-Inflammatory Cytokines in Autoantibody Positive First-Degree Relatives (FDRs) of Patients with Rheumatoid Arthritis. Hughes-Austin, J. M., Deane, K. D., Derber, L. A., Zerbe, G. O., Dabelea, D. M., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M. WILEY-BLACKWELL. 2012: S887

  View details for Web of Science ID 000309748305017

• Fine-Specificity of Anti-Citrullinated Peptide Auto-Antibodies: Associations with Cardiac Structure and Function in Rheumatoid Arthritis. Geraldino-Pardilla, L., Giles, J. T., Sokolove, J., Robinson, W. H., Bathon, J. M. WILEY-BLACKWELL. 2012: S1072

  View details for Web of Science ID 000309748306049

• An Expanded Repertoire of Anti-Citrullinated Peptide Antibodies Is Associated with Interstitial Lung Disease in Rheumatoid Arthritis Giles, J. T., Danoff, S., Sokolove, J., Winchester, R., Pappas, D. A., Cramb, C., Connors, G., Siegelman, S. S., Robinson, W. H., Bathon, J. M. WILEY-BLACKWELL. 2012: S1127

  View details for Web of Science ID 000309748306169

• Evolution of Preclinical Autoimmunity in Individuals At Risk for Development of Rheumatoid Arthritis Annual Scientific Meeting of the American-College-of-Rheumatology (ACR) and Association-of-Rheumatology-Health-Professionals (ARHP) El-Gabalawy, H. S., Robinson, D. B., Smolik, I., Hart, D. M., Ferucci, E. D., Newkirk, M. M., Fritzler, M. J., Cramb, C., Sokolove, J., Robinson, W. H. WILEY-BLACKWELL. 2012: S722–S722

  View details for Web of Science ID 000309748303409

• Anti-Citrullinated Peptide Autoantibodies to Rheumatoid Synovium Epitopes in Women and Risk of Future Rheumatoid Arthritis. Annual Scientific Meeting of the American-College-of-Rheumatology (ACR) and Association-of-Rheumatology-Health-Professionals (ARHP) Arkema, E. V., Goldstein, B. L., Robinson, W. H., Cramb, C., Sokolove, J., Cui, J., Malspeis, S., Karlson, E. W., Costenbader, K. H. WILEY-BLACKWELL. 2012: S685–S686

  View details for Web of Science ID 000309748303326

• Response to 'Plasma proteins present in osteoarthritic synovial fluid can stimulate cytokine production via Toll-like receptor 4' - authors' reply. Arthritis research & therapy Sohn, D. H., Sokolove, J., Sharpe, O., Erhart, J. C., Chandra, P. E., Lahey, L. J., Lindstrom, T. M., Hwang, I., Boyer, K. A., Andriacchi, T. P., Robinson, W. H. 2012; 14 (5): 406

  View details for DOI 10.1186/ar3892

  View details for PubMedID 22979902

  View details for PubMedCentralID PMC3580503

• Identification of Naturally Occurring Fatty Acids of the Myelin Sheath That Resolve Neuroinflammation SCIENCE TRANSLATIONAL MEDICINE Ho, P. P., Kanter, J. L., Johnson, A. M., Srinagesh, H. K., Chang, E., Purdy, T. M., Van Haren, K., Wikoff, W. R., Kind, T., Khademi, M., Matloff, L. Y., Narayana, S., Hur, E. M., Lindstrom, T. M., He, Z., Fiehn, O., Olsson, T., Han, X., Han, M. H., Steinman, L., Robinson, W. H. 2012; 4 (137)

  Abstract

  Lipids constitute 70% of the myelin sheath, and autoantibodies against lipids may contribute to the demyelination that characterizes multiple sclerosis (MS). We used lipid antigen microarrays and lipid mass spectrometry to identify bona fide lipid targets of the autoimmune response in MS brain, and an animal model of MS to explore the role of the identified lipids in autoimmune demyelination. We found that autoantibodies in MS target a phosphate group in phosphatidylserine and oxidized phosphatidylcholine derivatives. Administration of these lipids ameliorated experimental autoimmune encephalomyelitis by suppressing activation and inducing apoptosis of autoreactive T cells, effects mediated by the lipids' saturated fatty acid side chains. Thus, phospholipids represent a natural anti-inflammatory class of compounds that have potential as therapeutics for MS.

  View details for DOI 10.1126/scitranslmed.3003831

  View details for PubMedID 22674551

• New tools for classification and monitoring of autoimmune diseases NATURE REVIEWS RHEUMATOLOGY Maecker, H. T., Lindstrom, T. M., Robinson, W. H., Utz, P. J., Hale, M., Boyd, S. D., Shen-Orr, S. S., Fathman, C. G. 2012; 8 (6): 317-328

  Abstract

  Rheumatologists see patients with a range of autoimmune diseases. Phenotyping these diseases for diagnosis, prognosis and selection of therapies is an ever increasing problem. Advances in multiplexed assay technology at the gene, protein, and cellular level have enabled the identification of 'actionable biomarkers'; that is, biological metrics that can inform clinical practice. Not only will such biomarkers yield insight into the development, remission, and exacerbation of a disease, they will undoubtedly improve diagnostic sensitivity and accuracy of classification, and ultimately guide treatment. This Review provides an introduction to these powerful technologies that could promote the identification of actionable biomarkers, including mass cytometry, protein arrays, and immunoglobulin and T-cell receptor high-throughput sequencing. In our opinion, these technologies should become part of routine clinical practice for the management of autoimmune diseases. The use of analytical tools to deconvolve the data obtained from use of these technologies is also presented here. These analyses are revealing a more comprehensive and interconnected view of the immune system than ever before and should have an important role in directing future treatment approaches for autoimmune diseases.

  View details for DOI 10.1038/nrrheum.2012.66

  View details for PubMedID 22647780

• Autoantibody Epitope Spreading in the Pre-Clinical Phase Predicts Progression to Rheumatoid Arthritis PLOS ONE Sokolove, J., Bromberg, R., Deane, K. D., Lahey, L. J., Derber, L. A., Chandra, P. E., Edison, J. D., Gilliland, W. R., Tibshirani, R. J., Norris, J. M., Holers, V. M., Robinson, W. H. 2012; 7 (5)

  Abstract

  Rheumatoid arthritis (RA) is a prototypical autoimmune arthritis affecting nearly 1% of the world population and is a significant cause of worldwide disability. Though prior studies have demonstrated the appearance of RA-related autoantibodies years before the onset of clinical RA, the pattern of immunologic events preceding the development of RA remains unclear. To characterize the evolution of the autoantibody response in the preclinical phase of RA, we used a novel multiplex autoantigen array to evaluate development of the anti-citrullinated protein antibodies (ACPA) and to determine if epitope spread correlates with rise in serum cytokines and imminent onset of clinical RA. To do so, we utilized a cohort of 81 patients with clinical RA for whom stored serum was available from 1-12 years prior to disease onset. We evaluated the accumulation of ACPA subtypes over time and correlated this accumulation with elevations in serum cytokines. We then used logistic regression to identify a profile of biomarkers which predicts the imminent onset of clinical RA (defined as within 2 years of testing). We observed a time-dependent expansion of ACPA specificity with the number of ACPA subtypes. At the earliest timepoints, we found autoantibodies targeting several innate immune ligands including citrullinated histones, fibrinogen, and biglycan, thus providing insights into the earliest autoantigen targets and potential mechanisms underlying the onset and development of autoimmunity in RA. Additionally, expansion of the ACPA response strongly predicted elevations in many inflammatory cytokines including TNF-α, IL-6, IL-12p70, and IFN-γ. Thus, we observe that the preclinical phase of RA is characterized by an accumulation of multiple autoantibody specificities reflecting the process of epitope spread. Epitope expansion is closely correlated with the appearance of preclinical inflammation, and we identify a biomarker profile including autoantibodies and cytokines which predicts the imminent onset of clinical arthritis.

  View details for DOI 10.1371/journal.pone.0035296

  View details for PubMedID 22662108

• Pre-analytical effects of blood sampling and handling in quantitative immunoassays for rheumatoid arthritis JOURNAL OF IMMUNOLOGICAL METHODS Zhao, X., Qureshi, F., Eastman, P. S., Manning, W. C., Alexander, C., Robinson, W. H., Hesterberg, L. K. 2012; 378 (1-2): 72-80

  Abstract

  Variability in pre-analytical blood sampling and handling can significantly impact results obtained in quantitative immunoassays. Understanding the impact of these variables is critical for accurate quantification and validation of biomarker measurements. Particularly, in the design and execution of large clinical trials, even small differences in sample processing and handling can have dramatic effects in analytical reliability, results interpretation, trial management and outcome. The effects of two common blood sampling methods (serum vs. plasma) and two widely-used serum handling methods (on the clot with ambient temperature shipping, "traditional", vs. centrifuged with cold chain shipping, "protocol") on protein and autoantibody concentrations were examined. Matched serum and plasma samples were collected from 32 rheumatoid arthritis (RA) patients representing a wide range of disease activity status. Additionally, a set of matched serum samples with two sample handling methods was collected. One tube was processed per manufacturer's instructions and shipped overnight on cold packs (protocol). The matched tube, without prior centrifugation, was simultaneously shipped overnight at ambient temperatures (traditional). Upon delivery, the traditional tube was centrifuged. All samples were subsequently aliquoted and frozen prior to analysis of protein and autoantibody biomarkers. Median correlation between paired serum and plasma across all autoantibody assays was 0.99 (0.98-1.00) with a median % difference of -3.3 (-7.5 to 6.0). In contrast, observed protein biomarker concentrations were significantly affected by sample types, with median correlation of 0.99 (0.33-1.00) and a median % difference of -10 (-55 to 23). When the two serum collection/handling methods were compared, the median correlation between paired samples for autoantibodies was 0.99 (0.91-1.00) with a median difference of 4%. In contrast, significant increases were observed in protein biomarker concentrations among certain biomarkers in samples processed with the 'traditional' method. Autoantibody quantification appears robust to both sample type (plasma vs. serum) and pre-analytical sample collection/handling methods (protocol vs. traditional). In contrast, for non-antibody protein biomarker concentrations, sample type had a significant impact; plasma samples generally exhibit decreased protein biomarker concentrations relative to serum. Similarly, sample handling significantly impacted the variability of protein biomarker concentrations. When biomarker concentrations are combined algorithmically into a single test score such as a multi-biomarker disease activity test for rheumatoid arthritis (MBDA), changes in protein biomarker concentrations may result in a bias of the score. These results illustrate the importance of characterizing pre-analytical methodology, sample type, sample processing and handling procedures for clinical testing in order to ensure test accuracy.

  View details for DOI 10.1016/j.jim.2012.02.007

  View details for Web of Science ID 000304285900009

  View details for PubMedID 22366959

  View details for PubMedCentralID PMC3404505

• Inhibition of Epidermal Growth Factor Receptor Tyrosine Kinase Ameliorates Collagen-Induced Arthritis JOURNAL OF IMMUNOLOGY Swanson, C. D., Akama-Garren, E. H., Stein, E. A., Petralia, J. D., Ruiz, P. J., Edalati, A., Lindstrom, T. M., Robinson, W. H. 2012; 188 (7): 3513-3521

  Abstract

  Rheumatoid arthritis (RA) is an autoimmune synovitis characterized by the formation of pannus and the destruction of cartilage and bone in the synovial joints. Although immune cells, which infiltrate the pannus and promote inflammation, play a prominent role in the pathogenesis of RA, other cell types also contribute. Proliferation of synovial fibroblasts, for example, underlies the formation of the pannus, while proliferation of endothelial cells results in neovascularization, which supports the growth of the pannus by supplying it with nutrients and oxygen. The synovial fibroblasts also promote inflammation in the synovium by producing cytokines and chemokines. Finally, osteoclasts cause the destruction of bone. In this study, we show that erlotinib, an inhibitor of the tyrosine kinase epidermal growth factor receptor (EGFR), reduces the severity of established collagen-induced arthritis, a mouse model of RA, and that it does so by targeting synovial fibroblasts, endothelial cells, and osteoclasts. Erlotinib-induced attenuation of autoimmune arthritis was associated with a reduction in number of osteoclasts and blood vessels, and erlotinib inhibited the formation of murine osteoclasts and the proliferation of human endothelial cells in vitro. Erlotinib also inhibited the proliferation and cytokine production of human synovial fibroblasts in vitro. Moreover, EGFR was highly expressed and activated in the synovium of mice with collagen-induced arthritis and patients with RA. Taken together, these findings suggest that EGFR plays a central role in the pathogenesis of RA and that EGFR inhibition may provide benefits in the treatment of RA.

  View details for DOI 10.4049/jimmunol.1102693

  View details for PubMedID 22393153

• Therapeutic Effects of Systemic Administration of Chaperone alpha B-Crystallin Associated with Binding Proinflammatory Plasma Proteins JOURNAL OF BIOLOGICAL CHEMISTRY Rothbard, J. B., Kurnellas, M. P., Brownell, S., Adams, C. M., Su, L., Axtell, R. C., Chen, R., Fathman, C. G., Robinson, W. H., Steinman, L. 2012; 287 (13): 9708-9721

  Abstract

  The therapeutic benefit of the small heat shock protein αB-crystallin (HspB5) in animal models of multiple sclerosis and ischemia is proposed to arise from its increased capacity to bind proinflammatory proteins at the elevated temperatures within inflammatory foci. By mass spectral analysis, a common set of ∼70 ligands was precipitated by HspB5 from plasma from patients with multiple sclerosis, rheumatoid arthritis, and amyloidosis and mice with experimental allergic encephalomyelitis. These proteins were distinguished from other precipitated molecules because they were enriched in the precipitate as compared with their plasma concentrations, and they exhibited temperature-dependent binding. More than half of these ligands were acute phase proteins or members of the complement or coagulation cascades. Consistent with this proposal, plasma levels of HspB5 were increased in patients with multiple sclerosis as compared with normal individuals. The combination of the thermal sensitivity of the HspB5 combined with the high local concentration of these ligands at the site of inflammation is proposed to explain the paradox of how a protein believed to exhibit nonspecific binding can bind with some relative apparent selectivity to proinflammatory proteins and thereby modulate inflammation.

  View details for DOI 10.1074/jbc.M111.337691

  View details for PubMedID 22308023

• Differential mTOR and ERK pathway utilization by effector CD4 T cells suggests combinatorial drug therapy of arthritis CLINICAL IMMUNOLOGY Lin, J. T., Stein, E. A., Wong, M. T., Kalpathy, K. J., Su, L. L., Utz, P. J., Robinson, W. H., Fathnnan, C. G. 2012; 142 (2): 127-138

  Abstract

  The signaling pathways utilized by naïve and experienced effector CD4 T cells during activation and proliferation were evaluated. While inhibition of either mTOR or MAPK alone was able to inhibit naïve T cell proliferation, both mTOR and MAPK (ERK) pathway inhibition was required to efficiently block experienced, effector CD4 T cell proliferation. This was demonstrated both in vitro, and in vivo by treating mice with collagen-induced arthritis using mTOR and/or ERK inhibitors. The combination of mTOR and ERK inhibition prevented or treated disease more efficiently than either agent alone. These data illustrate the different requirements of naïve and experienced effector CD4 T cells in the use of the mTOR and MAPK pathways in proliferation, and suggest that therapies targeting both the mTOR and MAPK pathways may be more effective than targeting either pathway alone in the treatment of CD4 T cell-mediated autoimmunity.

  View details for DOI 10.1016/j.clim.2011.09.008

  View details for PubMedID 22075384

• Plasma proteins present in osteoarthritic synovial fluid can stimulate cytokine production via Toll-like receptor 4 ARTHRITIS RESEARCH & THERAPY Sohn, D. H., Sokolove, J., Sharpe, O., Erhart, J. C., Chandra, P. E., Lahey, L. J., Lindstrom, T. M., Hwang, I., Boyer, K. A., Andriacchi, T. P., Robinson, W. H. 2012; 14 (1)

  Abstract

  Osteoarthritis (OA) is a degenerative disease characterized by cartilage breakdown in the synovial joints. The presence of low-grade inflammation in OA joints is receiving increasing attention, with synovitis shown to be present even in the early stages of the disease. How the synovial inflammation arises is unclear, but proteins in the synovial fluid of affected joints could conceivably contribute. We therefore surveyed the proteins present in OA synovial fluid and assessed their immunostimulatory properties.We used mass spectrometry to survey the proteins present in the synovial fluid of patients with knee OA. We used a multiplex bead-based immunoassay to measure levels of inflammatory cytokines in serum and synovial fluid from patients with knee OA and from patients with rheumatoid arthritis (RA), as well as in sera from healthy individuals. Significant differences in cytokine levels between groups were determined by significance analysis of microarrays, and relations were determined by unsupervised hierarchic clustering. To assess the immunostimulatory properties of a subset of the identified proteins, we tested the proteins' ability to induce the production of inflammatory cytokines by macrophages. For proteins found to be stimulatory, the macrophage stimulation assays were repeated by using Toll-like receptor 4 (TLR4)-deficient macrophages.We identified 108 proteins in OA synovial fluid, including plasma proteins, serine protease inhibitors, proteins indicative of cartilage turnover, and proteins involved in inflammation and immunity. Multiplex cytokine analysis revealed that levels of several inflammatory cytokines were significantly higher in OA sera than in normal sera, and levels of inflammatory cytokines in synovial fluid and serum were, as expected, higher in RA samples than in OA samples. As much as 36% of the proteins identified in OA synovial fluid were plasma proteins. Testing a subset of these plasma proteins in macrophage stimulation assays, we found that Gc-globulin, α1-microglobulin, and α2-macroglobulin can signal via TLR4 to induce macrophage production of inflammatory cytokines implicated in OA.Our findings suggest that plasma proteins present in OA synovial fluid, whether through exudation from plasma or production by synovial tissues, could contribute to low-grade inflammation in OA by functioning as so-called damage-associated molecular patterns in the synovial joint.

  View details for DOI 10.1186/ar3555

  View details for Web of Science ID 000304698800021

  View details for PubMedID 22225630

  View details for PubMedCentralID PMC3392793

• Citrullination of fibrinogen: generation of neoepitopes and enhancement of immunostimulatory properties Robinson, W. H., Sokolove, J. BIOMED CENTRAL LTD. 2012

  View details for Web of Science ID 000303915700031

• Response to 'Plasma proteins present in osteoarthritic synovial fluid can stimulate cytokine production via Toll-like receptor 4' - authors' reply ARTHRITIS RESEARCH & THERAPY Sohn, D. H., Sokolove, J., Sharpe, O., Erhart, J. C., Chandra, P. E., Lahey, L. J., Lindstrom, T. M., Hwang, I., Boyer, K. A., Andriacchi, T. P., Robinson, W. H. 2012; 14 (5)

  View details for DOI 10.1186/ar3892

  View details for Web of Science ID 000315488700008

• Interleukin-34 produced by human fibroblast-like synovial cells in rheumatoid arthritis supports osteoclastogenesis ARTHRITIS RESEARCH & THERAPY Hwang, S., Choi, B., Kang, S., Chang, J., Kim, Y., Chung, Y., Sohn, D. H., So, M. W., Lee, C., Robinson, W. H., Chang, E. 2012; 14 (1)

  Abstract

  Interleukin-34 (IL-34) is a recently defined cytokine, showing a functional overlap with macrophage colony stimulating factor (M-CSF). This study was undertaken to address the expression of IL-34 in rheumatoid arthritis (RA) patients and to investigate its regulation and pathogenic role in RA.IL-34 levels were determined in the RA synovium, synovial fluid (SF) and fibroblast-like synovial cells (FLS) by immunohistochemistry, real-time PCR, enzyme-linked immunosorbent assay and immunoblotting. RA activity was assessed using Disease Activity Score 28 (DAS28) activity in the plasma collected at baseline and one year after treatment. Conditioned media (CM) were prepared from RA FLS culture with tumor necrosis factor alpha (TNFα) for 24 hours and used for functional assay.IL-34 was expressed in the synovium, SF, and FLS from RA patients. The production of IL-34 in FLS was up-regulated by TNFα in RA samples compared with osteoarthritis (OA) patients. Importantly, the preferential induction of IL-34 rather than M-CSF by TNFα in RAFLS was mediated by the transcription factor nuclear factor kappa B (NF-κB) and activation of c-Jun N-terminal kinase (JNK). IL-34 elevation in plasma from RA patients was decreased after the administration of disease-modifying anti-rheumatic drugs (DMARDs) in accordance with a decrease in DAS28. CM from RAFLS cultured with TNFα promoted chemotactic migration of human peripheral blood mononuclear cells (PBMCs) and subsequent osteoclast (OC) formation, effects that were attenuated by an anti-IL-34 antibody.These data provide novel information about the production of IL-34 in RA FLS and indicate that IL-34 is an additional osteoclastogenic factor regulated by TNFα in RA, suggesting a discrete role of IL-34 in inflammatory RA diseases.

  View details for DOI 10.1186/ar3693

  View details for Web of Science ID 000304698800028

  View details for PubMedID 22264405

  View details for PubMedCentralID PMC3392804

• Development and deployment of antigen arrays for investigation of B-cell fine specificity in autoimmune disease. Frontiers in bioscience (Elite edition) Sokolove, J., Lindstrom, T. M., Robinson, W. H. 2012; 4: 320-330

  Abstract

  Recent developments in proteomic technologies have enabled the high-throughput, multiplex measurement of large panels of antibodies in biological fluids of patients with immune-driven diseases. Antigen microarrays are increasingly being used to delineate the natural history of autoantibody formation and epitope spread, and thus gain insight into the pathogenesis of autoimmune diseases, as well as into host immunity and its shortcomings. Characterization of autoimmunity that precedes the onset of clinically apparent disease has the potential to guide disease prevention using either conventional immunosupression or novel, antigen-specific tolerizing therapies. In addition, autoantibody profiling has the potential to identify molecular subtypes of a disease, which could allow for prediction of disease outcomes such as severity, tissue damage, and response to therapy.

  View details for PubMedID 22201874

  View details for PubMedCentralID PMC3404510

• Tyrosine Kinase Inhibitors Ameliorate Autoimmune Encephalomyelitis in a Mouse Model of Multiple Sclerosis JOURNAL OF CLINICAL IMMUNOLOGY Crespo, O., Kang, S. C., Daneman, R., Lindstrom, T. M., Ho, P. P., Sobel, R. A., Steinman, L., Robinson, W. H. 2011; 31 (6): 1010-1020

  Abstract

  Multiple sclerosis is an autoimmune disease of the central nervous system characterized by neuroinflammation and demyelination. Although considered a T cell-mediated disease, multiple sclerosis involves the activation of both adaptive and innate immune cells, as well as resident cells of the central nervous system, which synergize in inducing inflammation and thereby demyelination. Differentiation, survival, and inflammatory functions of innate immune cells and of astrocytes of the central nervous system are regulated by tyrosine kinases. Here, we show that imatinib, sorafenib, and GW2580-small molecule tyrosine kinase inhibitors-can each prevent the development of disease and treat established disease in a mouse model of multiple sclerosis. In vitro, imatinib and sorafenib inhibited astrocyte proliferation mediated by the tyrosine kinase platelet-derived growth factor receptor (PDGFR), whereas GW2580 and sorafenib inhibited macrophage tumor necrosis factor (TNF) production mediated by the tyrosine kinases c-Fms and PDGFR, respectively. In vivo, amelioration of disease by GW2580 was associated with a reduction in the proportion of macrophages and T cells in the CNS infiltrate, as well as a reduction in the levels of circulating TNF. Our findings suggest that GW2580 and the FDA-approved drugs imatinib and sorafenib have potential as novel therapeutics for the treatment of autoimmune demyelinating disease.

  View details for DOI 10.1007/s10875-011-9579-6

  View details for PubMedID 21847523

• Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma JOURNAL OF BIOLOGICAL CHEMISTRY Zhao, L., Yamaguchi, Y., Sharif, S., Du, X., Song, J. J., Lee, D. M., Recht, L. D., Robinson, W. H., Morser, J., Leung, L. L. 2011; 286 (45): 39520-39527

  Abstract

  Chemerin is a chemoattractant involved in immunity that may also function as an adipokine. Chemerin circulates as an inactive precursor (chem163S), and its activation requires proteolytic cleavages at its C terminus, involving proteases involved in coagulation, fibrinolysis, and inflammation. However, the key proteolytic steps in prochemerin activation in vivo remain to be established. Previously, we have shown that C-terminal cleavage of chem163S by plasmin to chem158K, followed by a carboxypeptidase cleavage, leads to the most active isoform, chem157S. To identify and quantify the in vivo chemerin isoforms in biological specimens, we developed specific ELISAs for chem163S, chem158K, and chem157S, using antibodies raised against peptides from the C terminus of the different chemerin isoforms. We found that the mean plasma concentrations of chem163S, chem158K, and chem157S were 40 ± 7.9, 8.1 ± 2.9, and 0.7 ± 0.8 ng/ml, respectively. The total level of cleaved and noncleaved chemerins in cerebrospinal fluids was ∼10% of plasma levels whereas it was elevated ∼2-fold in synovial fluids from patients with arthritis. On the other hand, the fraction of cleaved chemerins was much higher in synovial fluid and cerebrospinal fluid samples than in plasma (∼75%, 50%, and 18% respectively). Chem158K was the dominant chemerin isoform, and it was not generated by ex vivo processing, indicating that cleavage of prochemerin at position Lys-158, whether by plasmin or another serine protease, represents a major step in prochemerin activation in vivo. Our study provides the first direct evidence that chemerin undergoes extensive proteolytic processing in vivo, underlining the importance of measuring individual isoforms.

  View details for DOI 10.1074/jbc.M111.258954

  View details for PubMedID 21930706

• Porphyromonas Gingivalis (P. gingivalis) is Associated with the Presence of Disease-Specific Autoantibodies in Individuals At Increased Risk for the Future Development of Rheumatoid Arthritis 75th Annual Scientific Meeting of the American-College-of-Rheumatology/46th Annual Scientific Meeting of the Association-of-Rheumatology-Health-Professionals (ARHP) Mikuls, T. R., Deane, K. D., Payne, J., Thiele, G. M., O'Dell, J. R., Derber, L. A., Robinson, W. H., Holers, V. M., Norris, J. M. WILEY-BLACKWELL. 2011: S300–S300

  View details for Web of Science ID 000297621500762

• The Association Between Rheumatoid Factor and Predisposing Factors for Atherogenesis in First-Degree Relatives without Rheumatoid Arthritis: Studies of the Etiology of Rheumatoid Arthritis Hughes-Austin, J. M., Deane, K. D., Derber, L. A., Zerbe, G. O., Dabelea, D. M., Eckel, R. H., Sokolove, J., Robinson, W. H., Holers, V., Norris, J. M. WILEY-BLACKWELL. 2011: S34

  View details for Web of Science ID 000297621500101

• Rheumatoid Arthritis Case Validation Strategies in Large Research Databases: The Women's Health Initiative Experience. Walitt, B. T., Mackey, R., Kuller, L., Deane, K. D., Robinson, W., Holers, V., Tracey, R., Chang, Y., Moreland, L. W. WILEY-BLACKWELL. 2011: S120–S121

  View details for Web of Science ID 000297621500322

• Serum Cytokine Levels Predict Mortality Among Postmenopausal Women Reporting Rheumatoid Arthritis. Sokolove, J., Robinson, W. H., Walitt, B. T., Deane, K. D., Chang, Y., Tracy, R., Holers, V., Kuller, L., Moreland, L. W., Mackey, R. WILEY-BLACKWELL. 2011: S144

  View details for Web of Science ID 000297621500388

• Development of a High-Throughput, Multiplex Assay for Profiling the Autoantibody Fine Specificity in Rheumatoid Arthritis Zhao, X., Eastman, P., Qureshi, F., Manning, W. C., Robinson, W., Hesterberg, L. K. WILEY-BLACKWELL. 2011: S686

  View details for Web of Science ID 000297621502134

• Smoking and Periodontal Status Are Associated with ACPA Fine Specificity and Levels of Inflammatory Cytokines in Rheumatoid Arthritis: The ARIC Study. Bromberg, R., Sokolove, J., Lahey, L. J., Alonso, A., Wener, M. H., Gersuk, V. H., Buckner, J., Beck, J. D., Michalowicz, B. S., Folsom, A. A., Robinson, W., Molitor, J. A. WILEY-BLACKWELL. 2011: S845–S846

  View details for Web of Science ID 000297621502556

• Plasma carboxypeptidase B downregulates inflammatory responses in autoimmune arthritis JOURNAL OF CLINICAL INVESTIGATION Song, J. J., Hwang, I., Cho, K. H., Garcia, M. A., Kim, A. J., Wang, T. H., Lindstrom, T. M., Lee, A. T., Nishimura, T., Zhao, L., Morser, J., Nesheim, M., Goodman, S. B., Lee, D. M., Bridges, S. L., Gregersen, P. K., Leung, L. L., Robinson, W. H. 2011; 121 (9): 3517-3527

  Abstract

  The immune and coagulation systems are both implicated in the pathogenesis of rheumatoid arthritis (RA). Plasma carboxypeptidase B (CPB), which is activated by the thrombin/thrombomodulin complex, plays a procoagulant role during fibrin clot formation. However, an antiinflammatory role for CPB is suggested by the recent observation that CPB can cleave proinflammatory mediators, such as C5a, bradykinin, and osteopontin. Here, we show that CPB plays a central role in downregulating C5a-mediated inflammatory responses in autoimmune arthritis. CPB deficiency exacerbated inflammatory arthritis in a mouse model of RA, and cleavage of C5a by CPB suppressed the ability of C5a to recruit immune cells in vivo. In human patients with RA, genotyping of nonsynonymous SNPs in the CPB-encoding gene revealed that the allele encoding a CPB variant with longer half-life was associated with a lower risk of developing radiographically severe RA. Functionally, this CPB variant was more effective at abrogating the proinflammatory properties of C5a. Additionally, expression of both CPB and C5a in synovial fluid was higher in patients with RA than in those with osteoarthritis. These findings suggest that CPB plays a critical role in dampening local, C5a-mediated inflammation and represents a molecular link between inflammation and coagulation in autoimmune arthritis.

  View details for DOI 10.1172/JCI46387

  View details for PubMedID 21804193

• Tyrosine kinases in inflammatory dermatologic disease JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY Paniagua, R. T., Fiorentino, D. F., Chung, L., Robinson, W. H. 2011; 65 (2): 389-403

  Abstract

  Tyrosine kinases (TKs) are enzymes that catalyze the phosphorylation of tyrosine residues on protein substrates. They are key components of signaling pathways that drive an array of cellular responses including proliferation, differentiation, migration, and survival. Specific TKs have recently been identified as critical to the pathogenesis of several autoimmune and inflammatory diseases. Small-molecule inhibitors of TKs are emerging as a novel class of therapy that may provide benefit in certain patient subsets. In this review, we highlight TK signaling implicated in inflammatory dermatologic diseases, evaluate strategies aimed at inhibiting these aberrant signaling pathways, and discuss prospects for future drug development.

  View details for DOI 10.1016/j.jaad.2010.04.026

  View details for PubMedID 20584561

• Role of the Toll-like receptor pathway in the recognition of orthopedic implant wear-debris particles BIOMATERIALS Pearl, J. I., Ma, T., Irani, A. R., Huang, Z., Robinson, W. H., Smith, R. L., Goodman, S. B. 2011; 32 (24): 5535-5542

  Abstract

  The inflammatory response to prosthetic implant-derived wear particles is the primary cause of bone loss and aseptic loosening of implants, but the mechanisms by which macrophages recognize and respond to particles remain unknown. Studies of innate immunity demonstrate that Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPS). All TLRs signal through myeloid differentiation factor 88 (MyD88), except TLR3 which signals through TIR domain containing adapter inducing interferon-beta (TRIF), and TLR4 which signals through both MyD88 and TRIF. We hypothesized that wear-debris particles may act as PAMPs/DAMPs and activate macrophages via TLRs. To test this hypothesis, we first demonstrated that inhibition of MyD88 decreases polymethylmethacrylate (PMMA) particle-induced production of TNF-α in RAW 264.7 macrophages. Next we compared particle-induced production of TNF-α among MyD88 knockout (MyD88(-/-)), TRIF knockout (TRIF(-/-)), and wild type (WT) murine macrophages. Relative to WT, disruption of MyD88 signaling diminished, and disruption of TRIF amplified the particle-induced production of TNF-α. Gene expression data indicated that this latter increase in TNF-α was due to a compensatory increase in expression of MyD88 associated components of the TLR pathway. Finally, using an in vivo model, MyD88(-/-) mice developed less particle-induced osteolysis than WT mice. These results indicate that the response to PMMA particles is partly dependent on MyD88, presumably as part of TLR signaling; MyD88 may represent a therapeutic target for prevention of wear debris-induced periprosthetic osteolysis.

  View details for DOI 10.1016/j.biomaterials.2011.04.046

  View details for PubMedID 21592562

• Human peptidome display NATURE BIOTECHNOLOGY Robinson, W. H., Steinman, L. 2011; 29 (6): 500-502

  View details for DOI 10.1038/nbt.1888

  View details for Web of Science ID 000291342100014

  View details for PubMedID 21654669

  View details for PubMedCentralID PMC3404725

• Chaperone Activity of alpha B-Crystallin Is Responsible for Its Incorrect Assignment as an Autoantigen in Multiple Sclerosis JOURNAL OF IMMUNOLOGY Rothbard, J. B., Zhao, X., Sharpe, O., Strohman, M. J., Kurnellas, M., Mellins, E. D., Robinson, W. H., Steinman, L. 2011; 186 (7): 4263-4268

  Abstract

  For 15 y, α B-crystallin (heat shock protein [Hsp] B5) has been labeled an autoantigen in multiple sclerosis (MS) based on humoral and cellular responses found in humans and animal models. However, there have been several scientific inconsistencies with this assignment, ranging from studies demonstrating small differences in anticrystallin responses between patients and healthy individuals to the inability of crystallin-specific T cells to induce symptoms of experimental allergic encephalomyelitis in animal models. Experiments in this article demonstrate that the putative anti-HspB5 Abs from 23 MS patients cross-react with 7 other members of the human small Hsp family and were equally present in normal plasma. Biolayer interferometry demonstrates that the binding was temperature dependent, and that the calculated K(a) increased as the concentration of the sHsp decreased. These two patterns are characteristic of multiple binding sites with varying affinities, the composition of which changes with temperature, supporting the hypothesis that HspB5 bound the Ab and not the reverse. HspB5 also precipitated Ig heavy and L chains from sera from patients with MS. These results establish that small Hsps bind Igs with high affinity and refute much of the serological data used to assign α B-crystallin as an autoantigen.

  View details for DOI 10.4049/jimmunol.1003934

  View details for PubMedID 21357544

• Development of peptide microarrays to investigate autoantibody epitope specificity in Pemphigus vulgaris 71st Annual Meeting of the Society-for-Investigative-Dermatology Hazelton, J., Patel, M., Furstenberg, G., Tyagi, S., Haab, B. B., Robinson, W. H., Seiffert-Sinha, K., Sinha, A. A. NATURE PUBLISHING GROUP. 2011: S9–S9

  View details for Web of Science ID 000289035600053

• N-alpha-Benzoyl-N5-(2-Chloro-1-Iminoethyl)-L-Ornithine Amide, a Protein Arginine Deiminase Inhibitor, Reduces the Severity of Murine Collagen-Induced Arthritis JOURNAL OF IMMUNOLOGY Willis, V. C., Gizinski, A. M., Banda, N. K., Causey, C. P., Knuckley, B., Cordova, K. N., Luo, Y., Levitt, B., Glogowska, M., Chandra, P., Kulik, L., Robinson, W. H., Arend, W. P., Thompson, P. R., Holers, V. M. 2011; 186 (7): 4396-4404

  Abstract

  Rheumatoid arthritis is associated with the development of autoantibodies to citrullinated self-proteins. Citrullinated synovial proteins, which are generated via the actions of the protein arginine deiminases (PADs), are known to develop in the murine collagen-induced arthritis (CIA) model of inflammatory arthritis. Given these findings, we evaluated whether N-α-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide (Cl-amidine), a recently described pan-PAD inhibitor, could affect the development of arthritis and autoimmunity by treating mice in the CIA model with Cl-amidine on days 0-35. Cl-amidine treatment reduced total synovial and serum citrullination, decreased clinical disease activity by ∼50%, and significantly decreased IgG2a anti-mouse type II collagen Abs. Additionally, histopathology scores and total complement C3 deposition were significantly lower in Cl-amidine-treated mice compared with vehicle controls. Synovial microarray analyses demonstrated decreased IgG reactivity to several native and citrullinated epitopes compared with vehicle controls. Cl-amidine treatment had no ameliorative effect on collagen Ab-induced arthritis, suggesting its primary protective mechanism was not mediated through effector pathways. Reduced levels of citrullinated synovial proteins observed in mice treated with Cl-amidine are consistent with the notion that Cl-amidine derives its efficacy from its ability to inhibit the deiminating activity of PADs. In total, these results suggested that PADs are necessary participants in the autoimmune and subsequent inflammatory processes in CIA. Cl-amidine may represent a novel class of disease-modifying agents that modulate aberrant citrullination, and perhaps other immune processes, necessary for the development of inflammatory arthritis.

  View details for DOI 10.4049/jimmunol.1001620

  View details for Web of Science ID 000288751200067

  View details for PubMedID 21346230

  View details for PubMedCentralID PMC3085980

• Fishing for Biomarkers with Antigen Mimics CELL Lindstrom, T. M., Robinson, W. H. 2011; 144 (1): 13-15

  Abstract

  Current efforts to identify antibodies that are biomarkers of disease rely on knowing the antigens they target. In many diseases, however, the relevant antigens are unknown. Reddy et al. (2010) now present an approach for discovering antibody biomarkers that avoids the need for antigen identification.

  View details for DOI 10.1016/j.cell.2010.12.022

  View details for PubMedID 21215365

• Immune Complexes Containing Citrullinated Fibrinogen Costimulate Macrophages via Toll-like Receptor 4 and Fc gamma Receptor ARTHRITIS AND RHEUMATISM Sokolove, J., Zhao, X., Chandra, P. E., Robinson, W. H. 2011; 63 (1): 53-62

  Abstract

  Rheumatoid arthritis (RA) is associated with the presence of anti-citrullinated protein antibodies (ACPAs). Nearly two-thirds of patients with ACPA-positive RA have immune complexes that contain citrullinated fibrinogen, and these citrullinated fibrinogen-containing immune complexes (cFb-IC) can exacerbate disease in murine models of RA; however, the exact role of such ACPA ICs in RA pathogenesis has remained elusive. We undertook the present study to investigate a novel mechanism by which ACPAs specifically targeting citrullinated fibrinogen may directly stimulate macrophage tumor necrosis factor (TNF) production.Murine or human macrophages were stimulated with native fibrinogen (nFb), cFb, or in vitro-generated nFb-IC or cFb-IC, and TNF production was measured by enzyme-linked immunosorbent assay. ICs were generated with either polyclonal anti-Fb antibodies or pooled IgG from patients with ACPA-positive RA. To evaluate the role of the Toll-like receptor 4 (TLR-4)/myeloid differentiation protein (MyD88) pathway and the Fcγ receptor (FcγR) pathway in the induction of TNF by Fb and Fb-IC, parallel experiments were performed using 1) TLR-4-deficient or MyD88-deficient macrophages, and 2) inhibitors of TLR-4 or FcγR.Citrullinated Fb stimulated macrophage TNF production more potently than did native Fb. Incorporation of cFb into ICs augmented its ability to stimulate TNF production by macrophages. Stimulation of TNF by cFb was dependent on TLR-4 and MyD88, while stimulation by cFb-IC was dependent on both TLR-4/MyD88 and FcγR.We demonstrated that cFb-IC can costimulate macrophages via dual engagement of TLR-4 and FcγR, resulting in the synergistic induction of TNF production. Our findings suggest a potential role of citrullination in increasing the potency of an endogenous innate immune ligand and provide insight into the mechanism by which anticitrulline autoimmunity may contribute to the onset and propagation of inflammation in RA.

  View details for DOI 10.1002/art.30081

  View details for PubMedID 20954191

• The Bruton tyrosine kinase inhibitor PCI-32765 ameliorates autoimmune arthritis by inhibition of multiple effector cells ARTHRITIS RESEARCH & THERAPY Chang, B. Y., Huang, M. M., Francesco, M., Chen, J., Sokolove, J., Magadala, P., Robinson, W. H., Buggy, J. J. 2011; 13 (4)

  Abstract

  The aim was to determine the effect of the Bruton tyrosine kinase (Btk)-selective inhibitor PCI-32765, currently in Phase I/II studies in lymphoma trials, in arthritis and immune-complex (IC) based animal models and describe the underlying cellular mechanisms.PCI-32765 was administered in a series of murine IC disease models including collagen-induced arthritis (CIA), collagen antibody-induced arthritis (CAIA), reversed passive anaphylactic reaction (RPA), and passive cutaneous anaphylaxis (PCA). Clinical and pathologic features characteristic of each model were examined following treatment. PCI-32765 was then examined in assays using immune cells relevant to the pathogenesis of arthritis, and where Btk is thought to play a functional role. These included proliferation and calcium mobilization in B cells, cytokine and chemokine production in monocytes/macrophages, degranulation of mast cells and its subsequent cytokine/chemokine production.PCI-32765 dose-dependently and potently reversed arthritic inflammation in a therapeutic CIA model with an ED(50) of 2.6 mg/kg/day. PCI-32765 also prevented clinical arthritis in CAIA models. In both models, infiltration of monocytes and macrophages into the synovium was completely inhibited and importantly, the bone and cartilage integrity of the joints were preserved. PCI-32765 reduced inflammation in the Arthus and PCA assays. In vitro, PCI-32765 inhibited BCR-activated primary B cell proliferation (IC(50) = 8 nM). Following FcγR stimulation, PCI-32765 inhibited TNFα, IL-1β and IL-6 production in primary monocytes (IC(50) = 2.6, 0.5, 3.9 nM, respectively). Following FcεRI stimulation of cultured human mast cells, PCI-32765 inhibited release of histamine, PGD(2), TNF-α, IL-8 and MCP-1.PCI-32765 is efficacious in CIA, and in IC models that do not depend upon autoantibody production from B cells. Thus PCI-32765 targets not only B lymphocytes but also monocytes, macrophages and mast cells, which are important Btk-expressing effector cells in arthritis.

  View details for DOI 10.1186/ar3400

  View details for Web of Science ID 000297150200006

  View details for PubMedID 21752263

  View details for PubMedCentralID PMC3239353

• Novel multiplex technology for diagnostic characterization of rheumatoid arthritis ARTHRITIS RESEARCH & THERAPY Chandra, P. E., Sokolove, J., Hipp, B. G., Lindstrom, T. M., Elder, J. T., Reveille, J. D., Eberl, H., Klause, U., Robinson, W. H. 2011; 13 (3)

  Abstract

  The aim of this study was to develop a clinical-grade, automated, multiplex system for the differential diagnosis and molecular stratification of rheumatoid arthritis (RA).We profiled autoantibodies, cytokines, and bone-turnover products in sera from 120 patients with a diagnosis of RA of < 6 months' duration, as well as in sera from 27 patients with ankylosing spondylitis, 28 patients with psoriatic arthritis, and 25 healthy individuals. We used a commercial bead assay to measure cytokine levels and developed an array assay based on novel multiplex technology (Immunological Multi-Parameter Chip Technology) to evaluate autoantibody reactivities and bone-turnover markers. Data were analyzed by Significance Analysis of Microarrays and hierarchical clustering software.We developed a highly reproducible, automated, multiplex biomarker assay that can reliably distinguish between RA patients and healthy individuals or patients with other inflammatory arthritides. Identification of distinct biomarker signatures enabled molecular stratification of early-stage RA into clinically relevant subtypes. In this initial study, multiplex measurement of a subset of the differentiating biomarkers provided high sensitivity and specificity in the diagnostic discrimination of RA: Use of 3 biomarkers yielded a sensitivity of 84.2% and a specificity of 93.8%, and use of 4 biomarkers a sensitivity of 59.2% and a specificity of 96.3%.The multiplex biomarker assay described herein has the potential to diagnose RA with greater sensitivity and specificity than do current clinical tests. Its ability to stratify RA patients in an automated and reproducible manner paves the way for the development of assays that can guide RA therapy.

  View details for DOI 10.1186/ar3383

  View details for PubMedID 21702928

• The Number of Elevated Cytokines and Chemokines in Preclinical Seropositive Rheumatoid Arthritis Predicts Time to Diagnosis in an Age-Dependent Manner ARTHRITIS AND RHEUMATISM Deane, K. D., O'Donnell, C. I., Hueber, W., Majka, D. S., Lazar, A. A., Derber, L. A., Gilliland, W. R., Edison, J. D., Norris, J. M., Robinson, W. H., Holers, V. M. 2010; 62 (11): 3161-3172

  Abstract

  To evaluate levels of biomarkers in preclinical rheumatoid arthritis (RA) and to use elevated biomarkers to develop a model for the prediction of time to future diagnosis of seropositive RA.Stored samples obtained from 73 military cases with seropositive RA prior to RA diagnosis and from controls (mean 2.9 samples per case; samples collected a mean of 6.6 years prior to diagnosis) were tested for rheumatoid factor (RF) isotypes, anti-cyclic citrullinated peptide (anti-CCP) antibodies, 14 cytokines and chemokines (by bead-based assay), and C-reactive protein (CRP).Preclinical positivity for anti-CCP and/or ≥2 RF isotypes was >96% specific for future RA. In preclinical RA, levels of the following were positive in a significantly greater proportion of RA cases versus controls: interleukin-1α (IL-1α), IL-1β, IL-6, IL-10, IL-12p40, IL-12p70, IL-15, fibroblast growth factor 2, flt-3 ligand, tumor necrosis factor α, interferon-γ-inducible 10-kd protein, granulocyte-macrophage colony-stimulating factor, and CRP. Also, increasing numbers of elevated cytokines/chemokines were present in cases nearer to the time of diagnosis. RA patients who were ≥40 years old at diagnosis had a higher proportion of samples positive for cytokines/chemokines 5-10 years prior to diagnosis than did patients who were <40 years old at diagnosis (P < 0.01). In regression modeling using only case samples positive for autoantibodies highly specific for future RA, increasing numbers of cytokines/chemokines were predictive of decreased time to diagnosis, and the predicted time to diagnosis based on cytokines/chemokines was longer in older compared with younger cases.Levels of autoantibodies, cytokines/chemokines, and CRP are elevated in the preclinical period of RA development. In preclinical autoantibody-positive cases, the number of elevated cytokines/chemokines is predictive of the time of diagnosis of future RA in an age-dependent manner.

  View details for DOI 10.1002/art.27638

  View details for Web of Science ID 000283776400005

  View details for PubMedID 20597112

  View details for PubMedCentralID PMC2980824

• A model for harmonizing flow cytometry in clinical trials NATURE IMMUNOLOGY Maecker, H. T., McCoy, J. P. 2010; 11 (11): 975-978

  Abstract

  Complexities in sample handling, instrument setup and data analysis are barriers to the effective use of flow cytometry to monitor immunological parameters in clinical trials. The novel use of a central laboratory may help mitigate these issues.

  View details for DOI 10.1038/ni1110-975

  View details for Web of Science ID 000283127800003

  View details for PubMedCentralID PMC3400260

• A model for harmonizing flow cytometry in clinical trials. Nature immunology Maecker, H. T., McCoy, J. P., Amos, M., Elliott, J., Gaigalas, A., Wang, L., Aranda, R., Banchereau, J., Boshoff, C., Braun, J., Korin, Y., Reed, E., Cho, J., Hafler, D., Davis, M., Fathman, C. G., Robinson, W., Denny, T., Weinhold, K., Desai, B., Diamond, B., Gregersen, P., Di Meglio, P., Nestle, F. O., Peakman, M., Villanova, F., Ferbas, J., Field, E., Kantor, A., Kawabata, T., Komocsar, W., Lotze, M., Nepom, J., Ochs, H., O'Lone, R., Phippard, D., Plevy, S., Rich, S., Roederer, M., Rotrosen, D., Yeh, J. 2010; 11 (11): 975-978

  Abstract

  Complexities in sample handling, instrument setup and data analysis are barriers to the effective use of flow cytometry to monitor immunological parameters in clinical trials. The novel use of a central laboratory may help mitigate these issues.

  View details for DOI 10.1038/ni1110-975

  View details for PubMedID 20959798

  View details for PubMedCentralID PMC3400260

• Evaluation of an imatinib response gene signature in patients with systemic sclerosis Chung, L., Ruiz, P., Wood, T., Shoor, S., Robinson, W., Whitfield, M., Chang, H., Fiorentino, D. CLINICAL & EXPER RHEUMATOLOGY. 2010: S62–S62

  View details for Web of Science ID 000284028600028

• Rheumatoid Arthritis: A Role for Immunosenescence? JOURNAL OF THE AMERICAN GERIATRICS SOCIETY Lindstrom, T. M., Robinson, W. H. 2010; 58 (8): 1565-1575

  Abstract

  Aging is accompanied by a progressive decline in the integrity of the immune system, a process known as immunosenescence. Pathological features typical of immune dysfunction in older adults, encompassing dysregulation of innate and adaptive immune responses, characterize rheumatoid arthritis (RA), an autoimmune disease whose incidence increases with age. Recent evidence suggests that certain features of immunosenescence, such as the decrease in T-cell generation and diversity, may contribute to the development of RA. Thus, physiological immunosenescence may render older adults susceptible to RA, and premature immunosenescence may contribute to the development of RA in young adults. In addition, other features of immunosenescence may result from the chronic immune stimulation that occurs in RA and lead to worsening of the disease. This article reviews the immunopathological features common to aging and RA and discusses the mechanisms by which immunosenescence may contribute to the development or progression of RA.

  View details for DOI 10.1111/j.1532-5415.2010.02965.x

  View details for Web of Science ID 000280643700021

  View details for PubMedID 20942872

  View details for PubMedCentralID PMC3055796

• Natural killer cells trigger osteoclastogenesis and bone destruction in arthritis PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Soederstroem, K., Stein, E., Colmenero, P., Purath, U., Mueller-Ladner, U., de Matos, C. T., Tarner, I. H., Robinson, W. H., Engleman, E. G. 2010; 107 (29): 13028-13033

  Abstract

  Osteoclasts are bone-eroding cells that develop from monocytic precursor cells in the presence of receptor activator of NF-kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Osteoclasts are essential for physiological bone remodeling, but localized excessive osteoclast activity is responsible for the periarticular bone destruction that characteristically occurs in patients with rheumatoid arthritis (RA). The origin of osteoclasts at sites of bone erosion in RA is unknown. Natural killer (NK) cells, as well as monocytes, are abundant in the inflamed joints of patients with RA. We show here that such NK cells express both RANKL and M-CSF and are frequently associated with CD14(+) monocytes in the RA synovium. Moreover, when synovial NK cells are cocultured with monocytes in vitro, they trigger their differentiation into osteoclasts, a process dependent on RANKL and M-CSF. As in RA, NK cells in the joints of mice with collagen-induced arthritis (CIA) express RANKL. Depletion of NK cells from mice before the induction of CIA reduces the severity of subsequent arthritis and almost completely prevents bone erosion. These results suggest that NK cells may play an important role in the destruction of bone associated with inflammatory arthritis.

  View details for DOI 10.1073/pnas.1000546107

  View details for PubMedID 20615964

• Biomarkers for rheumatoid arthritis: making it personal. Scandinavian journal of clinical and laboratory investigation. Supplementum Lindstrom, T. M., Robinson, W. H. 2010; 242: 79-84

  Abstract

  Effective treatment of rheumatoid arthritis (RA) has been hampered by the heterogeneity of the disease. Although early intervention can result in disease remission, it requires early diagnosis - and current diagnostic tests are not sufficiently accurate or sensitive in the early stages of RA. As a result, RA is typically diagnosed only once damage to the joints has already begun, a time at which the window for optimal treatment may have been missed. Furthermore, a significant proportion of RA patients do not respond to any given therapeutic. Research efforts are increasingly focused on discovery of biomarkers that enable early diagnosis and stratification of RA, and thus the implementation of timely, targeted therapy. Biomarkers have the potential to transform the management of RA by enabling not only early diagnosis, but also assessment and prediction of disease severity, selection of therapy, and monitoring of response to therapy. In this mini review, we discuss the development of molecular biomarkers for RA.

  View details for DOI 10.3109/00365513.2010.493406

  View details for PubMedID 20515283

• Biomarkers for rheumatoid arthritis: Making it personal SCANDINAVIAN JOURNAL OF CLINICAL & LABORATORY INVESTIGATION Lindstrom, T. M., Robinson, W. H. 2010; 70: 79-84

  Abstract

  Effective treatment of rheumatoid arthritis (RA) has been hampered by the heterogeneity of the disease. Although early intervention can result in disease remission, it requires early diagnosis - and current diagnostic tests are not sufficiently accurate or sensitive in the early stages of RA. As a result, RA is typically diagnosed only once damage to the joints has already begun, a time at which the window for optimal treatment may have been missed. Furthermore, a significant proportion of RA patients do not respond to any given therapeutic. Research efforts are increasingly focused on discovery of biomarkers that enable early diagnosis and stratification of RA, and thus the implementation of timely, targeted therapy. Biomarkers have the potential to transform the management of RA by enabling not only early diagnosis, but also assessment and prediction of disease severity, selection of therapy, and monitoring of response to therapy. In this mini review, we discuss the development of molecular biomarkers for RA.

  View details for DOI 10.3109/00365513.2010.493406

  View details for Web of Science ID 000280333600017

  View details for PubMedCentralID PMC3417774

• Endogenous antibodies promote rapid myelin clearance and effective axon regeneration after nerve injury PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Vargas, M. E., Watanabe, J., Singh, S. J., Robinson, W. H., Barres, B. A. 2010; 107 (26): 11993-11998

  Abstract

  Degenerating myelin inhibits axon regeneration and is rapidly cleared after peripheral (PNS) but not central nervous system (CNS) injury. To better understand mechanisms underlying rapid PNS myelin clearance, we tested the potential role of the humoral immune system. Here, we show that endogenous antibodies are required for rapid and robust PNS myelin clearance and axon regeneration. B-cell knockout JHD mice display a significant delay in macrophage influx, myelin clearance, and axon regeneration. Rapid clearance of myelin debris is restored in mutant JHD mice by passive transfer of antibodies from naïve WT mice or by an anti-PNS myelin antibody, but not by delivery of nonneural antibodies. We demonstrate that degenerating nerve tissue is targeted by preexisting endogenous antibodies that control myelin clearance by promoting macrophage entrance and phagocytic activity. These results demonstrate a role for immunoglobulin (Ig) in clearing damaged self during healing and suggest that the immune-privileged status of the CNS may contribute to failure of CNS myelin clearance and axon regeneration after injury.

  View details for DOI 10.1073/pnas.1001948107

  View details for PubMedID 20547838

• A Multitude of Kinases-Which are the Best Targets in Treating Rheumatoid Arthritis? RHEUMATIC DISEASE CLINICS OF NORTH AMERICA Lindstrom, T. M., Robinson, W. H. 2010; 36 (2): 367-?

  Abstract

  Small-molecule kinase inhibitors are increasingly taking center stage in the quest for new drugs for the treatment of rheumatoid arthritis (RA). By targeting kinases, small-molecule inhibitors can exert potent anti-inflammatory and immunomodulatory effects; the success of small-molecule kinase inhibitors in the treatment of cancer has spurred efforts to identify kinases that could be targeted for the treatment of chronic inflammatory disorders, such as RA. Although many kinase inhibitors have proved efficacious in the treatment of inflammatory arthritis in animals few have been tested in RA clinical trials. This article discusses the challenges and progress in the pursuit of small-molecule kinase inhibitors for RA, including lessons learned from the failure of erstwhile frontrunner inhibitors and the promise of inhibitors making their debut on the RA stage.

  View details for DOI 10.1016/j.rdc.2010.02.005

  View details for Web of Science ID 000279254800010

  View details for PubMedID 20510239

  View details for PubMedCentralID PMC2879404

• Development of protein microarrays to investigate autoantibody profiles in pemphigus vulgaris Annual Meeting of the Society-for-Investigative-Dermatology Patel, M., Furstenberg, G., Hazelton, J., Tiagi, S., Seiffert-Sinha, K., Haab, B. B., Robinson, W. H., Sinha, A. A. NATURE PUBLISHING GROUP. 2010: S19–S19

  View details for Web of Science ID 000276455100111

• Autoimmunity against Fibrinogen Mediates Inflammatory Arthritis in Mice JOURNAL OF IMMUNOLOGY Ho, P. P., Lee, L. Y., Zhao, X., Tomooka, B. H., Paniagua, R. T., Sharpe, O., Benbarak, M. J., Chandra, P. E., Hueber, W., Steinman, L., Robinson, W. H. 2010; 184 (1): 379-390

  Abstract

  Rheumatoid arthritis (RA) is an autoimmune synovitis characterized by the presence of anticitrullinated protein Abs, although the exact targets and role of anticitrullinated protein autoimmunity in the pathogenesis of RA remain to be defined. Fibrinogen, which can be citrullinated, has recently emerged as a candidate autoantigen. To determine whether autoimmunity against fibrinogen can mediate inflammatory arthritis, we immunized a variety of common mouse strains with fibrinogen and found that DBA/1 and SJL mice developed an inflammatory and erosive arthritis. Mice with fibrinogen-induced arthritis (FIA) possess fibrinogen-reactive T cells that produce the proinflammatory cytokines IL-6, IL-17, TNF-alpha, and IFN-gamma. FIA can be adoptively transferred with either plasma or fibrinogen-specific T cells from diseased mice. Mice with FIA possess rheumatoid factor, circulating immune complexes, and anticyclic citrullinated peptide Abs, all of which are characteristic of human RA. These observations demonstrate that fibrinogen is arthritogenic in mice and that the pathogenesis of FIA is mediated by both autoantibodies and fibrinogen-reactive T cells.

  View details for DOI 10.4049/jimmunol.0901639

  View details for PubMedID 19949094

• Citrullinated Fibrinogen Stimulates TNF Release via TLR4 and Citrullinated Fibrinogen Immune Complexes Co-stimulate through TLR4 and the Fc Gamma Receptor Sokolove, J., Zhao, X., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2010: S7

  View details for DOI 10.1016/j.clim.2010.03.028

  View details for Web of Science ID 000277953700012

• Citrullinated Fibrinogen Stimulates TNF Release via TLR4 and Citrullinated Fibrinogen Immune Complexes Co-stimulate Through TLR4 and the Fc Gamma Receptor Sokolove, J., Zhao, X., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2010: S38–S39

  View details for DOI 10.1016/j.clim.2010.03.117

  View details for Web of Science ID 000277953700101

• EGFR Inhibition Ameliorates Murine Collagen-induced Arthritis Swanson, C., Robinson, W. H. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2010: S59

  View details for DOI 10.1016/j.clim.2010.03.181

  View details for Web of Science ID 000277953700165

• Correlation between Circulating Plasmablasts and Disease Activity in Patients with ACPA plus Rheumatoid Arthritis 10th Annual Meeting of the Federation-of-Clinical-Immunology-Societies Tan, Y. C., Sokolove, J., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2010: S138–S138

  View details for DOI 10.1016/j.clim.2010.03.418

  View details for Web of Science ID 000277953700402

• c-Fms-mediated differentiation and priming of monocyte lineage cells play a central role in autoimmune arthritis ARTHRITIS RESEARCH & THERAPY Paniagua, R. T., Chang, A., Mariano, M. M., Stein, E. A., Wang, Q., Lindstrom, T. M., Sharpe, O., Roscow, C., Ho, P. P., Lee, D. M., Robinson, W. H. 2010; 12 (1)

  Abstract

  Tyrosine kinases are key mediators of multiple signaling pathways implicated in rheumatoid arthritis (RA). We previously demonstrated that imatinib mesylate--a Food and Drug Administration (FDA)-approved, antineoplastic drug that potently inhibits the tyrosine kinases Abl, c-Kit, platelet-derived growth factor receptor (PDGFR), and c-Fms--ameliorates murine autoimmune arthritis. However, which of the imatinib-targeted kinases is the principal culprit in disease pathogenesis remains unknown. Here we examine the role of c-Fms in autoimmune arthritis.We tested the therapeutic efficacy of orally administered imatinib or GW2580, a small molecule that specifically inhibits c-Fms, in three mouse models of RA: collagen-induced arthritis (CIA), anti-collagen antibody-induced arthritis (CAIA), and K/BxN serum transfer-induced arthritis (K/BxN). Efficacy was evaluated by visual scoring of arthritis severity, paw thickness measurements, and histological analysis. We assessed the in vivo effects of imatinib and GW2580 on macrophage infiltration of synovial joints in CIA, and their in vitro effects on macrophage and osteoclast differentiation, and on osteoclast-mediated bone resorption. Further, we determined the effects of imatinib and GW2580 on the ability of macrophage colony-stimulating factor (M-CSF; the ligand for c-Fms) to prime bone marrow-derived macrophages to produce tumor necrosis factor (TNF) upon subsequent Fc receptor ligation. Finally, we measured M-CSF levels in synovial fluid from patients with RA, osteoarthritis (OA), or psoriatic arthritis (PsA), and levels of total and phosphorylated c-Fms in synovial tissue from patients with RA.GW2580 was as efficacious as imatinib in reducing arthritis severity in CIA, CAIA, and K/BxN models of RA. Specific inhibition of c-Fms abrogated (i) infiltration of macrophages into synovial joints of arthritic mice; (ii) differentiation of monocytes into macrophages and osteoclasts; (iii) osteoclast-mediated bone resorption; and (iv) priming of macrophages to produce TNF upon Fc receptor stimulation, an important trigger of synovitis in RA. Expression and activation of c-Fms in RA synovium were high, and levels of M-CSF were higher in RA synovial fluid than in OA or PsA synovial fluid.These results suggest that c-Fms plays a central role in the pathogenesis of RA by mediating the differentiation and priming of monocyte lineage cells. Therapeutic targeting of c-Fms could provide benefit in RA.

  View details for DOI 10.1186/ar2940

  View details for PubMedID 20181277

• Targeting mTOR and MAPK Pathways to Inhibit Naive and Experienced Effector CD4 T Cells in Autoimmunity 10th Annual Meeting of the Federation-of-Clinical-Immunology-Societies Lin, J., Stein, E., Wong, M., Kalpathy, K., Su, L., Utz, P., Robinson, W., Fathman, C. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2010: S69–S69

  View details for DOI 10.1016/j.clim.2010.03.211

  View details for Web of Science ID 000277953700195

• Autoimmune Disease Classification by Inverse Association with SNP Alleles PLOS GENETICS Sirota, M., Schaub, M. A., Batzoglou, S., Robinson, W. H., Butte, A. J. 2009; 5 (12)

  Abstract

  With multiple genome-wide association studies (GWAS) performed across autoimmune diseases, there is a great opportunity to study the homogeneity of genetic architectures across autoimmune disease. Previous approaches have been limited in the scope of their analysis and have failed to properly incorporate the direction of allele-specific disease associations for SNPs. In this work, we refine the notion of a genetic variation profile for a given disease to capture strength of association with multiple SNPs in an allele-specific fashion. We apply this method to compare genetic variation profiles of six autoimmune diseases: multiple sclerosis (MS), ankylosing spondylitis (AS), autoimmune thyroid disease (ATD), rheumatoid arthritis (RA), Crohn's disease (CD), and type 1 diabetes (T1D), as well as five non-autoimmune diseases. We quantify pair-wise relationships between these diseases and find two broad clusters of autoimmune disease where SNPs that make an individual susceptible to one class of autoimmune disease also protect from diseases in the other autoimmune class. We find that RA and AS form one such class, and MS and ATD another. We identify specific SNPs and genes with opposite risk profiles for these two classes. We furthermore explore individual SNPs that play an important role in defining similarities and differences between disease pairs. We present a novel, systematic, cross-platform approach to identify allele-specific relationships between disease pairs based on genetic variation as well as the individual SNPs which drive the relationships. While recognizing similarities between diseases might lead to identifying novel treatment options, detecting differences between diseases previously thought to be similar may point to key novel disease-specific genes and pathways.

  View details for DOI 10.1371/journal.pgen.1000792

  View details for PubMedID 20041220

• Thrombin-Activatable Carboxypeptidase B Cleavage of Osteopontin Regulates Neutrophil Survival and Synoviocyte Binding in Rheumatoid Arthritis ARTHRITIS AND RHEUMATISM Sharif, S. A., Du, X., Myles, T., Song, J. J., Price, E., Lee, D. M., Goodman, S. B., Nagashima, M., Morser, J., Robinson, W. H., Leung, L. L. 2009; 60 (10): 2902-2912

  Abstract

  Osteopontin (OPN) is a proinflammatory cytokine that plays an important role in the pathogenesis of rheumatoid arthritis (RA). OPN can be cleaved by thrombin, resulting in OPN-R and exposing the cryptic C-terminal alpha4beta1 and alpha9beta1 integrin-binding motif (SVVYGLR). Thrombin-activatable carboxypeptidase B (CPB), also called thrombin-activatable fibrinolysis inhibitor, removes the C-terminal arginine from OPN-R, generating OPN-L and abrogating its enhanced cell binding. We undertook this study to investigate the roles of OPN-R and OPN-L in synoviocyte adhesion, which contributes to the formation of invasive pannus, and in neutrophil survival, which affects inflammatory infiltrates in RA.Using specifically developed enzyme-linked immunosorbent assays, we tested the synovial fluid of patients with RA, osteoarthritis (OA), and psoriatic arthritis (PsA) to determine OPN-R, OPN-L, and full-length OPN (OPN-FL) levels.Elevated levels of OPN-R and OPN-L were found in synovial fluid samples from RA patients, but not in samples from OA or PsA patients. Increased levels of OPN-R and OPN-L correlated with increased levels of multiple inflammatory cytokines, including tumor necrosis factor alpha and interleukin-6. Immunohistochemical analyses revealed robust expression of OPN-FL, but only minimal expression of OPN-R, in RA synovium, suggesting that cleaved OPN is released into synovial fluid. In cellular assays, OPN-FL, and to a lesser extent OPN-R and OPN-L, had an antiapoptotic effect on neutrophils. OPN-R augmented RA fibroblast-like synoviocyte binding mediated by SVVYGLR binding to alpha4beta1, whereas OPN-L did not.Thrombin activation of OPN (resulting in OPN-R) and its subsequent inactivation by thrombin-activatable CPB (generating OPN-L) occurs locally within inflamed joints in RA. Our data suggest that thrombin-activatable CPB plays a central homeostatic role in RA by regulating neutrophil viability and reducing synoviocyte adhesion.

  View details for DOI 10.1002/art.24814

  View details for PubMedID 19790060

• A broad screen for targets of immune complexes decorating arthritic joints highlights deposition of nucleosomes in rheumatoid arthritis PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Monach, P. A., Hueber, W., Kessler, B., Tomooka, B. H., BenBarak, M., Simmons, B. P., Wright, J., Thornhill, T. S., Monestier, M., Ploegh, H., Robinson, W. H., Mathis, D., Benoist, C. 2009; 106 (37): 15867-15872

  Abstract

  Deposits of Ig and complement are abundant in affected joints of patients with rheumatoid arthritis (RA) and in animal models of RA in which antibodies are demonstrably pathogenic. To identify molecular targets of the Igs deposited in arthritic joints, which may activate local inflammation, we used a combination of mass spectrometry (MS) and protein microarrays. Immune complexes were affinity-purified from surgically removed joint tissues of 26 RA and osteoarthritis (OA) patients. Proteins complexed with IgG were identified by proteomic analysis using tandem MS. A striking diversity of components of the extracellular matrix, and some intracellular components, copurified specifically with IgG from RA and OA tissues. A smaller set of autoantigens was observed only in RA eluates. In complementary experiments, IgG fractions purified from joint immune complexes were tested on protein microarrays against a range of candidate autoantigens. These Igs bound a diverse subset of proteins and peptides from synovium and cartilage, different from that bound by normal serum Ig. One type of intracellular protein detected specifically in RA joints (histones H2A/B) was validated by immunohistology and found to be deposited on the cartilage surface of RA but not OA joints. Thus, autoantibodies to many determinants (whether deposited as "neoantigens" or normal constituents of the extracellular matrix) have the potential to contribute to arthritic inflammation.

  View details for DOI 10.1073/pnas.0908032106

  View details for Web of Science ID 000269806600065

  View details for PubMedID 19720992

  View details for PubMedCentralID PMC2747210

• Engagement of CD81 induces ezrin tyrosine phosphorylation and its cellular redistribution with filamentous actin JOURNAL OF CELL SCIENCE Coffey, G. P., Rajapaksa, R., Liu, R., Sharpe, O., Kuo, C., Krauss, S. W., Sagi, Y., Davis, R. E., Staudt, L. M., Sharman, J. P., Robinson, W. H., Levy, S. 2009; 122 (17): 3137-3144

  Abstract

  CD81 is a tetraspanin family member involved in diverse cellular interactions in the immune and nervous systems and in cell fusion events. However, the mechanism of action of CD81 and of other tetraspanins has not been defined. We reasoned that identifying signaling molecules downstream of CD81 would provide mechanistic clues. We engaged CD81 on the surface of B-lymphocytes and identified the induced tyrosine-phosphorylated proteins by mass spectrometry. This analysis showed that the most prominent tyrosine phosphorylated protein was ezrin, an actin-binding protein and a member of the ezrin-radixin-moesin family. We also found that CD81 engagement induces spleen tyrosine kinase (Syk) and that Syk was involved in tyrosine phosphorylation of ezrin. After engagement of CD81, it colocalized with ezrin and F-actin, and this association was disrupted when Syk activation was blocked. Taken together, these studies suggest a model in which CD81 interfaces between the plasma membrane and the cytoskeleton by activating Syk, mobilizing ezrin, and recruiting F-actin to facilitate cytoskeletal reorganization and cell signaling. This mechanism might explain the pleiotropic effects induced in response to stimulation of cells by anti-CD81 antibodies or by the hepatitis C virus, which uses this molecule as its key receptor.

  View details for DOI 10.1242/jcs.045658

  View details for PubMedID 19654214

• Neuroprotective natural antibodies to assemblies of amyloidogenic peptides decrease with normal aging and advancing Alzheimer's disease PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Britschgi, M., Olin, C. E., Johns, H. T., Takeda-Uchimura, Y., LeMieux, M. C., Rufibach, K., Rajadas, J., Zhang, H., Tomooka, B., Robinson, W. H., Clark, C. M., Fagan, A. M., Galasko, D. R., Holtzman, D. M., Jutel, M., Kaye, J. A., Lemere, C. A., Leszek, J., Li, G., Peskind, E. R., Quinn, J. F., Yesavage, J. A., Ghiso, J. A., Wyss-Coray, T. 2009; 106 (29): 12145-12150

  Abstract

  A number of distinct beta-amyloid (Abeta) variants or multimers have been implicated in Alzheimer's disease (AD), and antibodies recognizing such peptides are in clinical trials. Humans have natural Abeta-specific antibodies, but their diversity, abundance, and function in the general population remain largely unknown. Here, we demonstrate with peptide microarrays the presence of natural antibodies against known toxic Abeta and amyloidogenic non-Abeta species in plasma samples and cerebrospinal fluid of AD patients and healthy controls aged 21-89 years. Antibody reactivity was most prominent against oligomeric assemblies of Abeta and pyroglutamate or oxidized residues, and IgGs specific for oligomeric preparations of Abeta1-42 in particular declined with age and advancing AD. Most individuals showed unexpected antibody reactivities against peptides unique to autosomal dominant forms of dementia (mutant Abeta, ABri, ADan) and IgGs isolated from plasma of AD patients or healthy controls protected primary neurons from Abeta toxicity. Aged vervets showed similar patterns of plasma IgG antibodies against amyloid peptides, and after immunization with Abeta the monkeys developed high titers not only against Abeta peptides but also against ABri and ADan peptides. Our findings support the concept of conformation-specific, cross-reactive antibodies that may protect against amyloidogenic toxic peptides. If a therapeutic benefit of Abeta antibodies can be confirmed in AD patients, stimulating the production of such neuroprotective antibodies or passively administering them to the elderly population may provide a preventive measure toward AD.

  View details for DOI 10.1073/pnas.0904866106

  View details for Web of Science ID 000268178400059

  View details for PubMedID 19581601

  View details for PubMedCentralID PMC2715538

• Tyrosine kinases as targets for the treatment of rheumatoid arthritis NATURE REVIEWS RHEUMATOLOGY Swanson, C. D., Paniagua, R. T., Lindstrom, T. M., Robinson, W. H. 2009; 5 (6): 317-324

  Abstract

  As critical regulators of numerous cell signaling pathways, tyrosine kinases are implicated in the pathogenesis of several diseases, including rheumatoid arthritis (RA). In the absence of disease, synoviocytes produce factors that provide nutrition and lubrication for the surrounding cartilage tissue; few cellular infiltrates are seen in the synovium. In RA, however, macrophages, neutrophils, T cells and B cells infiltrate the synovium and produce cytokines, chemokines and degradative enzymes that promote inflammation and joint destruction. In addition, the synovial lining expands owing to the proliferation of synoviocytes and infiltration of inflammatory cells to form a pannus, which invades the surrounding bone and cartilage. Many of these cell responses are regulated by tyrosine kinases that operate in specific signaling pathways, and inhibition of a number of these kinases might be expected to provide benefit in RA.

  View details for DOI 10.1038/nrrheum.2009.82

  View details for PubMedID 19491913

• Identification of mRNA binding proteins that regulate the stability of LDL receptor mRNA through AU-rich elements JOURNAL OF LIPID RESEARCH Li, H., Chen, W., Zhou, Y., Abidi, P., Sharpe, O., Robinson, W. H., Kraemer, F. B., Liu, J. 2009; 50 (5): 820-831

  Abstract

  The 3'untranslated region (UTR) of human LDL receptor (LDLR) mRNA contains three AU-rich elements (AREs) responsible for rapid mRNA turnover and mediates the stabilization induced by berberine (BBR). However, the identities of the specific RNA binding proteins involved in the regulation of LDLR mRNA stability at the steady state level or upon BBR treatment are unknown. By conducting small interfering RNA library screenings, biotinylated RNA pull-down, mass spectrometry analysis, and functional assays, we now identify heterogeneous nuclear ribonucleoprotein D (hnRNP D), hnRNP I, and KH-type splicing regulatory protein (KSRP) as key modulators of LDLR mRNA stability in liver cells. We show that hnRNP D, I, and KSRP interact with AREs of the LDLR 3'UTR with sequence specificity. Silencing the expression of these proteins increased LDLR mRNA and protein levels. We further demonstrate that BBR-induced mRNA stabilization involves hnRNP I and KSRP, as their cellular depletions abolished the BBR effect and BBR treatment reduced the binding of hnRNP I and KSRP to the LDLR mRNA 3'UTR. These new findings demonstrate that LDLR mRNA stability is controlled by a group of ARE binding proteins, including hnRNP D, hnRNP I, and KSRP. Our results suggest that interference with the ability of destabilizing ARE binding proteins to interact with LDLR-ARE motifs is likely a mechanism for regulating LDLR expression by compounds such as BBR and perhaps others.

  View details for DOI 10.1194/jlr.M800375-JLR200

  View details for Web of Science ID 000264969300006

  View details for PubMedID 19141871

  View details for PubMedCentralID PMC2666168

• Elemental bio-imaging of calcium phosphate crystal deposits in knee samples from arthritic patients METALLOMICS Austin, C., Hare, D., Rozelle, A. L., Robinson, W. H., Grimm, R., Doble, P. 2009; 1 (2): 142-147

  Abstract

  Laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS) was employed to image deposits of calcium phosphate based crystals in knee cartilage and synovial fluid from arthritic patients. A reaction/collision cell containing hydrogen minimised plasma interferences on calcium and also improved the image quality without significant sensitivity reduction. Areas of high calcium and phosphorus intensities consistent with crystal deposits were observed for both the cartilage and synovial fluid samples. These areas were also characterised by high magnesium and strontium intensities. Distribution patterns of other elements such as copper and sulfur did not correlate with the crystal deposits. Filtered and non-filtered solutions of calcium phosphate crystals grown in synthetic synovial fluid were also imaged as further evidence of crystal deposits. The crystal deposits were detected in the unfiltered solution, and were absent from the filtered solutions.

  View details for DOI 10.1039/b901310p

  View details for Web of Science ID 000269034100003

  View details for PubMedID 21305107

  View details for PubMedCentralID PMC3405194

• Use of imatinib to treat systemic sclerosis: A prospective case series Arefiev, K., Fiorentino, D., Chung, L., Robinson, W. MOSBY-ELSEVIER. 2009: AB3

  View details for Web of Science ID 000263934100010

• Molecular Framework for Response to Imatinib Mesylate in Systemic Sclerosis ARTHRITIS AND RHEUMATISM Chung, L., Fiorentino, D. F., Benbarak, M. J., Adler, A. S., Mariano, M. M., Paniagua, R. T., Milano, A., Connolly, M. K., Ratiner, B. D., Wiskocil, R. L., Whitfield, M. L., Chang, H. Y., Robinson, W. H. 2009; 60 (2): 584-591

  Abstract

  Systemic sclerosis (SSc) is an autoimmune disease in which the tyrosine kinases platelet-derived growth factor receptor (PDGFR) and Abl are hypothesized to contribute to the fibrosis and vasculopathy of the skin and internal organs. Herein we describe 2 patients with early diffuse cutaneous SSc (dcSSc) who experienced reductions in cutaneous sclerosis in response to therapy with the tyrosine kinase inhibitor imatinib mesylate. Immunohistochemical analyses of skin biopsy specimens demonstrated reductions of phosphorylated PDGFRbeta and Abl with imatinib therapy. By gene expression profiling, an imatinib-responsive signature specific to dcSSc was identified (P < 10(-8)). The response of these patients and the findings of the analyses suggest that PDGFRbeta and Abl play critical, synergistic roles in the pathogenesis of SSc, and that imatinib targets a gene expression program that is frequently dysregulated in dcSSc.

  View details for DOI 10.1002/art.24221

  View details for PubMedID 19180499

• Blood autoantibody and cytokine profiles predict response to anti-tumor necrosis factor therapy in rheumatoid arthritis ARTHRITIS RESEARCH & THERAPY Hueber, W., Tomooka, B. H., Batliwalla, F., Li, W., Monach, P. A., Tibshirani, R. J., Van Vollenhoven, R. F., Lampa, J., Saito, K., Tanaka, Y., Genovese, M. C., Klareskog, L., Gregersen, P. K., Robinson, W. H. 2009; 11 (3)

  Abstract

  Anti-TNF therapies have revolutionized the treatment of rheumatoid arthritis (RA), a common systemic autoimmune disease involving destruction of the synovial joints. However, in the practice of rheumatology approximately one-third of patients demonstrate no clinical improvement in response to treatment with anti-TNF therapies, while another third demonstrate a partial response, and one-third an excellent and sustained response. Since no clinical or laboratory tests are available to predict response to anti-TNF therapies, great need exists for predictive biomarkers.Here we present a multi-step proteomics approach using arthritis antigen arrays, a multiplex cytokine assay, and conventional ELISA, with the objective to identify a biomarker signature in three ethnically diverse cohorts of RA patients treated with the anti-TNF therapy etanercept.We identified a 24-biomarker signature that enabled prediction of a positive clinical response to etanercept in all three cohorts (positive predictive values 58 to 72%; negative predictive values 63 to 78%).We identified a multi-parameter protein biomarker that enables pretreatment classification and prediction of etanercept responders, and tested this biomarker using three independent cohorts of RA patients. Although further validation in prospective and larger cohorts is needed, our observations demonstrate that multiplex characterization of autoantibodies and cytokines provides clinical utility for predicting response to the anti-TNF therapy etanercept in RA patients.

  View details for DOI 10.1186/ar2706

  View details for PubMedID 19460157

• Cardiac Glycosides as Anti-inflammatory Agents in Collagen Arthritis Stein, E., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2009: S74

  View details for DOI 10.1016/j.clim.2009.03.216

  View details for Web of Science ID 000266342300209

• B Cells and Transplantation: An Educational Resource Tandem Meeting on Biology of Blood and Marrow Transplantation Small, T. N., Robinson, W. H., Miklos, D. B. ELSEVIER SCIENCE INC. 2009: 104–13

  View details for DOI 10.1016/j.bbmt.2008.10.016

  View details for Web of Science ID 000263034300022

  View details for PubMedID 19147088

  View details for PubMedCentralID PMC2944827

• Tyrosine Kinases in Autoimmune Demyelination 9th Annual Meeting of the Federation-of-Clinical-Immunology-Societies Crespo, O., Kang, S., Daneman, R., Sobel, R., Ho, P., Steinman, L., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2009: S73–S73

  View details for DOI 10.1016/j.clim.2009.03.212

  View details for Web of Science ID 000266342300205

• Genomics and proteomics: Applications in autoimmune diseases. Pharmacogenomics and personalized medicine Hueber, W., Robinson, W. H. 2009; 2: 39-48

  Abstract

  Tremendous progress has been made over the past decade in the development and refinement of genomic and proteomic technologies for the identification of novel drug targets and molecular signatures associated with clinically important disease states, disease subsets, or differential responses to therapies. The rapid progress in high-throughput technologies has been preceded and paralleled by the elucidation of cytokine networks, followed by the stepwise clinical development of pathway-specific biological therapies that revolutionized the treatment of autoimmune diseases. Together, these advances provide opportunities for a long-anticipated personalized medicine approach to the treatment of autoimmune disease. The ever-increasing numbers of novel, innovative therapies will need to be harnessed wisely to achieve optimal long-term outcomes in as many patients as possible while complying with the demands of health authorities and health care providers for evidence-based, economically sound prescription of these expensive drugs. Genomic and proteomic profiling of patients with autoimmune diseases holds great promise in two major clinical areas: (1) rapid identification of new targets for the development of innovative therapies and (2) identification of patients who will experience optimal benefit and minimal risk from a specific (targeted) therapy. In this review, we attempt to capture important recent developments in the application of genomic and proteomic technologies to translational research by discussing informative examples covering a diversity of autoimmune diseases.

  View details for PubMedID 23226033

• Differentially Cleaved Forms of Osteopontin by Thrombin and Carboxypeptidase B (TAF1a) Regulates Neutrophil Viability and Synoviocyte Binding in Rheumatoid Arthritis 50th Annual Meeting of the American-Society-of-Hematology/ASH/ASCO Joint Symposium Sharif, S., Du, X., Myles, T., Song, J. J., Price, E. A., Lee, D., Goodman, S., Morser, J., Robinson, W., Leung, L. L. AMER SOC HEMATOLOGY. 2008: 1219–19

  View details for Web of Science ID 000262104704180

• Identification of acute phase reactants and cytokines useful for monitoring infliximab therapy in ankylosing spondylitis CLINICAL RHEUMATOLOGY Romero-Sanchez, C., Robinson, W. H., Tomooka, B. H., Londono, J., Valle-Onate, R., Huang, F., Deng, X., Zhang, L., Yang, C., Yu, D. T. 2008; 27 (11): 1429-1435

  Abstract

  Although most ankylosing spondylitis patients show an apparent clinical response to infliximab therapy, there is considerable individual variation. Because current clinical assessment relies heavily on subjective patient self-evaluation, biomarkers of high sensitivity and specificity are much needed. Here, we assessed potential biomarkers in 47 ankylosing spondylitis patients who received three standard pulses of infliximab. Before each infusion and at week 10, the following were measured: erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), platelet count, serum levels of metalloproteinase-3 (MMP-3), and 22 different cytokines. We discovered that, 2 weeks after the first infusion, the combination of ESR, CRP, and platelet count distinguished responders from non-responders with 81.3% sensitivity and 72.7% specificity. The distinguishing power was much less when each acute phase reactant was used alone. Among the 22 cytokines, serum IL-1alpha was able to distinguish responders from non-responders at week 6, with sensitivity of 84.9% and specificity of 53.8%. Serum IL-1alpha was probably generated from the joint compartments, as synovial fluid levels were much higher than corresponding serum levels. Although infliximab infusions led to rapid and significant suppression of serum MMP-3 levels, serum MMP-3 levels did not distinguish responders from non-responders. Besides identifying potential biomarkers, our results also demonstrate the usefulness of using sensitivity and specificity to assess usefulness of potential biomarkers.

  View details for DOI 10.1007/s10067-008-0941-x

  View details for Web of Science ID 000259816200013

  View details for PubMedID 18566849

• Regulation of tissue inflammation by thrombi n-activatable carboxypeptidase B (or TAFI) 22nd International Complement Workshop Leung, L. L., Myles, T., Nishimura, T., Song, J. J., Robinson, W. H. PERGAMON-ELSEVIER SCIENCE LTD. 2008: 4080–83

  Abstract

  Thrombin-activatable procarboxypeptidase B (proCPB or thrombin-activatable fibrinolysis inhibitor or TAFI) is a plasma procarboxypeptidase that is activated by the thrombin-thrombomodulin complex on the vascular endothelial surface. The activated CPB removes the newly exposed carboxyl terminal lysines in the partially digested fibrin clot, diminishes tissue plasminogen activator and plasminogen binding, and protects the clot from premature lysis. We have recently shown that CPB is catalytically more efficient than plasma CPN, the major plasma anaphylatoxin inhibitor, in inhibiting bradykinin, activated complement C3a, C5a, and thrombin-cleaved osteopontin in vitro. Using a thrombin mutant (E229K) that has minimal procoagulant properties but retains the ability to activate protein C and proCPB in vivo, we showed that infusion of E229K thrombin into wild-type mice reduced bradykinin-induced hypotension but it had no effect in proCPB-deficient mice, indicating that the beneficial effect of E229K thrombin is mediated through its activation of proCPB and not protein C. Similarly proCPB-deficient mice displayed enhanced pulmonary inflammation in a C5a-induced alveolitis model and E229K thrombin ameliorated the magnitude of alveolitis in wild-type but not proCPB-deficient mice. ProCPB-deficient mice also displayed enhanced arthritis in an inflammatory arthritis model. Thus, our in vitro and in vivo data support the thesis that thrombin-activatable CPB has broad anti-inflammatory properties. By specific cleavage of the carboxyl terminal arginines from C3a, C5a, bradykinin and thrombin-cleaved osteopontin, it inactivates these active inflammatory mediators. Along with the activation of protein C, the activation of proCPB by the endothelial thrombin-thrombomodulin complex represents a homeostatic feedback mechanism in regulating thrombin's pro-inflammatory functions in vivo.

  View details for DOI 10.1016/j.molimm.2008.07.010

  View details for PubMedID 18706698

• Anti-GITR treatment in collagen-induced arthritis results in enhanced epitope spreading and increased antibodies to citrullinated protein antigens (ACPA) 72nd Annual Scientific Meeting of the American-College-of-Rheumatology/43rd Annual Scientific Meeting of the Association-of-Rheumatology-Health-Professionals Cozine, C., Robinson, W. H., BenBarak, M., Holers, V. M. WILEY-BLACKWELL. 2008: S405–S406

  View details for Web of Science ID 000259244200669

• Phase 2 follow-up results of the BHT-3009 DNA vaccine for multiple sclerosis Havrdova, E., Krasulova, E., Nadj, C. G., Selmaj, K., Losy, J., Nadj, I., Radue, E., Gianettoni, J., Valone, F., Utz, P. J., Robinson, W. H., Steinman, L., Garren, H., BHT 3009 Study Grp SAGE PUBLICATIONS LTD. 2008: S47

  View details for Web of Science ID 000259675700138

• Cerebrospinal fluid cell phenotypic analysis from a phase 2 trial of the BHT-3009 DNA vaccine for multiple sclerosis Nadj, C., Nadj, I., Havrdova, E., Krasulova, E., Selmaj, K., Losy, J., Radue, E., Gianettoni, J., Solvason, N., Tersini, K., Valone, F., Utz, P. J., Robinson, W. H., Steinman, L., Garren, H., BHT 3009 Study Grp SAGE PUBLICATIONS LTD. 2008: S54

  View details for Web of Science ID 000259675700161

• Lipids as autoantigens and therapeutic immune modulators in multiple sclerosis Ho, P. R., Kanter, J. L., Johnson, A. M., Steinman, L., Robinson, W. H. SAGE PUBLICATIONS LTD. 2008: S13

  View details for Web of Science ID 000259675700031

• Rheumatoid arthritis (RA)-related antibody positivity is associated with serum cytokine elevations in RA-free first-degree relatives (FDRs) of RA-probands 72nd Annual Scientific Meeting of the American-College-of-Rheumatology/43rd Annual Scientific Meeting of the Association-of-Rheumatology-Health-Professionals Deane, K. D., Robinson, W. H., Majka, D. S., Parrish, L. A., BenBarak, M., Weisman, M. H., O'Dell, J. R., Gregersen, P. K., Buckner, J. H., Norris, J. M., Holers, V. M. WILEY-BLACKWELL. 2008: S615–S615

  View details for Web of Science ID 000259244201458

• A central anti-inflammatory role of carboxypeptidase B in autoimmune arthritis 72nd Annual Scientific Meeting of the American-College-of-Rheumatology/43rd Annual Scientific Meeting of the Association-of-Rheumatology-Health-Professionals Song, J. J., Nishimura, T., Garcia, M., Myles, T., Du, X., Sharif, S., Leung, L., Robinson, W. WILEY-BLACKWELL. 2008: S264–S265

  View details for Web of Science ID 000259244200285

• Molecular framework for response to imatinib mesylate in systemic sclerosis 72nd Annual Scientific Meeting of the American-College-of-Rheumatology/43rd Annual Scientific Meeting of the Association-of-Rheumatology-Health-Professionals Chung, L., Fiorentino, D. F., Benbarak, M. J., Adler, A. S., Mariano, M. M., Paniagua, R. T., Milano, A., Connolly, M. K., Ratiner, B. D., Wiskocil, R. L., Whitfield, M. L., Chang, H. Y., Robinson, W. H. WILEY-BLACKWELL. 2008: S819–S820

  View details for Web of Science ID 000259244202263

• Genomic and proteomic analysis reveals a threshold level of MYC required for tumor maintenance CANCER RESEARCH Shachaf, C. M., Gentles, A. J., Elchuri, S., Sahoo, D., Soen, Y., Sharpe, O., Perez, O. D., Chang, M., Mitchel, D., Robinson, W. H., Dill, D., Nolan, G. P., Plevritis, S. K., Felsher, D. W. 2008; 68 (13): 5132-5142

  Abstract

  MYC overexpression has been implicated in the pathogenesis of most types of human cancers. MYC is likely to contribute to tumorigenesis by its effects on global gene expression. Previously, we have shown that the loss of MYC overexpression is sufficient to reverse tumorigenesis. Here, we show that there is a precise threshold level of MYC expression required for maintaining the tumor phenotype, whereupon there is a switch from a gene expression program of proliferation to a state of proliferative arrest and apoptosis. Oligonucleotide microarray analysis and quantitative PCR were used to identify changes in expression in 3,921 genes, of which 2,348 were down-regulated and 1,573 were up-regulated. Critical changes in gene expression occurred at or near the MYC threshold, including genes implicated in the regulation of the G(1)-S and G(2)-M cell cycle checkpoints and death receptor/apoptosis signaling. Using two-dimensional protein analysis followed by mass spectrometry, phospho-flow fluorescence-activated cell sorting, and antibody arrays, we also identified changes at the protein level that contributed to MYC-dependent tumor regression. Proteins involved in mRNA translation decreased below threshold levels of MYC. Thus, at the MYC threshold, there is a loss of its ability to maintain tumorigenesis, with associated shifts in gene and protein expression that reestablish cell cycle checkpoints, halt protein translation, and promote apoptosis.

  View details for DOI 10.1158/0008-5472.CAN-07-6192

  View details for PubMedID 18593912

• Complement receptor CR2/CR1 deficiency protects mice from collagen-induced arthritis and associates with reduced autoantibodies to type II collagen and citrullinated antigens MOLECULAR IMMUNOLOGY Kuhn, K. A., Cozine, C. L., Tomooka, B., Robinson, W. H., Holers, V. M. 2008; 45 (10): 2808-2819

  Abstract

  Collagen-induced arthritis (CIA), a model of autoimmune inflammatory arthritis, depends upon complement activation and effective B cell responses. To determine the importance of complement receptors CR2/CR1 in CIA, the Cr2-/- genotype was backcrossed onto the DBA/1j strain. CIA was induced by immunization with bovine type II collagen in CFA on days 0 and 21. Cr2-/- mice demonstrated a significantly diminished arthritis severity, decreased antibodies to bovine and murine collagen, and a significant reduction in antibodies to citrullinated antigens. Autoantibodies to citrullinated antigens have been shown to amplify anti-type II collagen passive transfer arthritis. To test the hypothesis that that simple replacement of such antibodies might re-establish severe disease in Cr2-/- mice, monoclonal antibodies to citrullinated antigens were administered to mice during the disease course. Although citrullinated antigens targeted by these antibodies were present within the joints of all mice, addition of these monoclonal antibodies increased disease severity only in Cr2+/+ mice. Taken together, these data suggest that CR2/CR1 are required to develop robust autoimmunity in the CIA model and that amplification of arthritis by antibodies to citrullinated antigens depends on factor(s) absent in arthritic Cr2-/- mice.

  View details for DOI 10.1016/j.molimm.2008.01.036

  View details for Web of Science ID 000256318800011

  View details for PubMedID 18374982

• Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets NATURE Han, M. H., Hwang, S., Roy, D. B., Lundgren, D. H., Price, J. V., Ousman, S. S., Fernald, G. H., Gerlitz, B., Robinson, W. H., Baranzini, S. E., Grinnell, B. W., Raine, C. S., Sobel, R. A., Han, D. K., Steinman, L. 2008; 451 (7182): 1076-U2

  Abstract

  Understanding the neuropathology of multiple sclerosis (MS) is essential for improved therapies. Therefore, identification of targets specific to pathological types of MS may have therapeutic benefits. Here we identify, by laser-capture microdissection and proteomics, proteins unique to three major types of MS lesions: acute plaque, chronic active plaque and chronic plaque. Comparative proteomic profiles identified tissue factor and protein C inhibitor within chronic active plaque samples, suggesting dysregulation of molecules associated with coagulation. In vivo administration of hirudin or recombinant activated protein C reduced disease severity in experimental autoimmune encephalomyelitis and suppressed Th1 and Th17 cytokines in astrocytes and immune cells. Administration of mutant forms of recombinant activated protein C showed that both its anticoagulant and its signalling functions were essential for optimal amelioration of experimental autoimmune encephalomyelitis. A proteomic approach illuminated potential therapeutic targets selective for specific pathological stages of MS and implicated participation of the coagulation cascade.

  View details for DOI 10.1038/nature06559

  View details for PubMedID 18278032

• Epitope spreading to citrullinated antigens in mouse models of autoimmune arthritis and demyelination ARTHRITIS RESEARCH & THERAPY Kidd, B. A., Ho, P. P., Sharpe, O., Zhao, X., Tomooka, B. H., Kanter, J. L., Steinman, L., Robinson, W. H. 2008; 10 (5)

  Abstract

  Anti-citrullinated protein antibodies have a diagnostic role in rheumatoid arthritis (RA); however, little is known about their origins and contribution to pathogenesis. Citrullination is the post-translational conversion of arginine to citrulline by peptidyl arginine deiminase, and increased citrullination of proteins is observed in the joint tissue in RA and in brain tissue in multiple sclerosis (MS).We applied synovial and myelin protein arrays to examine epitope spreading of B cell responses to citrullinated epitopes in both the collagen-induced arthritis (CIA) model for RA and the experimental autoimmune encephalomyelitis (EAE) model for MS. Synovial and myelin protein arrays contain a spectrum of proteins and peptides, including native and citrullinated forms, representing candidate autoantigens in RA and MS, respectively. We applied these arrays to characterise the specificity of autoantibodies in serial serum samples derived from mice with acute and chronic stages of CIA and EAE.In samples from pre-disease CIA and acute-disease EAE, we observed autoantibody targeting of the immunising antigen and responses to a limited set of citrullinated epitopes. Over the course of diseases, the autoantibody responses expanded to target multiple citrullinated epitopes in both CIA and EAE. Using immunoblotting and mass spectrometry analysis, we identified citrullination of multiple polypeptides in CIA joint and EAE brain tissue that have not previously been described as citrullinated.Our results suggest that anti-citrulline antibody responses develop in the early stages of CIA and EAE, and that autoimmune inflammation results in citrullination of joint proteins in CIA and brain proteins in EAE, thereby creating neoantigens that become additional targets in epitope spreading of autoimmune responses.

  View details for DOI 10.1186/ar2523

  View details for PubMedID 18826638

• Lipids as autoantigens and therapeutic immune modulators in autoimmune demyelination Ho, P., Kanter, J., Johnson, A., Steinman, L., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2008: S24–S25

  View details for DOI 10.1016/j.clim.2008.03.064

  View details for Web of Science ID 000255533200065

• Induction of antigen-specific tolerance in multiple sclerosis after immunization with DNA encoding myelin basic protein in a randomized, placebo-controlled phase 1/2 trial ARCHIVES OF NEUROLOGY Bar-Or, A., Vollmer, T., Antel, J., Arnold, D. L., Bodner, C. A., Campagnolo, D., Gianettoni, J., Jalili, F., Kachuck, N., Lapierre, Y., Niino, M., Oger, J., Price, M., Rhodes, S., Robinson, W. H., Shi, F., Utz, P. J., Valone, F., Weiner, L., Steinman, L., Garren, H. 2007; 64 (10): 1407-1415

  Abstract

  To assess safety and immune modulation by BHT-3009, a tolerizing DNA vaccine encoding full-length human myelin basic protein, in patients with multiple sclerosis (MS).The study was a randomized, double-blind, placebo-controlled trial. Subjects receiving placebo were crossed over into an active arm after treatment unblinding.The trial was conducted at 4 academic institutions within North America. Patients Thirty patients with relapsing-remitting or secondary progressive MS who were not taking any other disease-modifying drugs were enrolled in the trial. Further, the patients were required to have either 1 to 5 gadolinium-enhancing lesions on screening brain magnetic resonance imaging (MRI), a relapse in the previous 2 years, or disease worsening in the previous 2 years.BHT-3009 was administered as intramuscular injections at weeks 1, 3, 5, and 9 after randomization into the trial, with or without 80 mg of daily oral atorvastatin calcium in combination. Three dose levels of BHT-3009 were tested (0.5 mg, 1.5 mg, and 3 mg).The primary outcome measures were safety and tolerability of BHT-3009. Secondary outcome measures included the number and volume of gadolinium-enhanced lesions on MRI, relapses, and analysis of antigen-specific immune responses.BHT-3009 was safe and well tolerated, provided favorable trends on brain MRI, and produced beneficial antigen-specific immune changes. These immune changes consisted of a marked decrease in proliferation of interferon-gamma-producing, myelin-reactive CD4+ T cells from peripheral blood and a reduction in titers of myelin-specific autoantibodies from cerebral spinal fluid as assessed by protein microarrays. We did not observe a substantial benefit of the atorvastatin combination compared with BHT-3009 alone.In patients with MS, BHT-3009 is safe and induces antigen-specific immune tolerance with concordant reduction of inflammatory lesions on brain MRI.

  View details for Web of Science ID 000249998400004

  View details for PubMedID 17698695

• Protective and therapeutic role for alpha B-crystallin in autoimmune demyelination NATURE Ousman, S. S., Tomooka, B. H., Van Noort, J. M., Wawrousek, E. F., O'Connor, K. C., Hafler, D. A., Sobel, R. A., Robinson, W. H., Steinman, L. 2007; 448 (7152): 474-U7

  Abstract

  alphaB-crystallin (CRYAB) is the most abundant gene transcript present in early active multiple sclerosis lesions, whereas such transcripts are absent in normal brain tissue. This crystallin has anti-apoptotic and neuroprotective functions. CRYAB is the major target of CD4+ T-cell immunity to the myelin sheath from multiple sclerosis brain. The pathophysiological implications of this immune response were investigated here. We demonstrate that CRYAB is a potent negative regulator acting as a brake on several inflammatory pathways in both the immune system and central nervous system (CNS). Cryab-/- mice showed worse experimental autoimmune encephalomyelitis (EAE) at the acute and progressive phases, with higher Th1 and Th17 cytokine secretion from T cells and macrophages, and more intense CNS inflammation, compared with their wild-type counterparts. Furthermore, Cryab-/- astrocytes showed more cleaved caspase-3 and more TUNEL staining, indicating an anti-apoptotic function of Cryab. Antibody to CRYAB was detected in cerebrospinal fluid from multiple sclerosis patients and in sera from mice with EAE. Administration of recombinant CRYAB ameliorated EAE. Thus, the immune response against a negative regulator of inflammation, CRYAB, in multiple sclerosis, would exacerbate inflammation and demyelination. This can be countered by giving CRYAB itself for therapy of ongoing disease.

  View details for DOI 10.1038/nature05935

  View details for PubMedID 17568699

• Proteomic analysis of secreted proteins in early rheumatoid arthritis: anti-citrulline autoreactivity is associated with up regulation of proinflammatory cytokines ANNALS OF THE RHEUMATIC DISEASES Hueber, W., Tomooka, B. H., Zhao, X., Kidd, B. A., Drijfhout, J. W., Fries, J. F., van Venrooij, W. J., Metzger, A. L., Genovese, M. C., Robinson, W. H. 2007; 66 (6): 712-719

  Abstract

  To identify peripheral blood autoantibody and cytokine profiles that characterise clinically relevant subgroups of patients with early rheumatoid arthritis using arthritis antigen microarrays and a multiplex cytokine assay.Serum samples from 56 patients with a diagnosis of rheumatoid arthritis of <6 months' duration were tested. Cytokine profiles were also determined in samples from patients with psoriatic arthritis (PsA) and ankylosing spondylitis (n = 21), and from healthy individuals (n = 19). Data were analysed using Kruskal-Wallis test with Dunn's adjustment for multiple comparisons, linear correlation tests, significance analysis of microarrays (SAM) and hierarchical clustering software.Distinct antibody profiles were associated with subgroups of patients who exhibited high serum levels of tumour necrosis factor (TNF)alpha, interleukin (IL)1beta, IL6, IL13, IL15 and granulocyte macrophage colony-stimulating factor. Significantly increased autoantibody reactivity against citrullinated epitopes was observed in patients within the cytokine "high" subgroup. Increased levels of TNFalpha, IL1alpha, IL12p40 and IL13, and the chemokines eotaxin/CCL11, monocyte chemoattractant protein-1 and interferon-inducible protein 10, were present in early rheumatoid arthritis as compared with controls (p<0.001). Chemokines showed some of the most impressive differences. Only IL8/CXCL8 concentrations were higher in patients with PsA/ankylosing spondylitis (p = 0.02).Increased blood levels of proinflammatory cytokines are associated with autoantibody targeting of citrullinated antigens and surrogate markers of disease activity in patients with early rheumatoid arthritis. Proteomic analysis of serum autoantibodies, cytokines and chemokines enables stratification of patients with early rheumatoid arthritis into molecular subgroups.

  View details for DOI 10.1136/ard.2006.054924

  View details for PubMedID 16901957

• Identification of biomarkers associated with the development of hepatocellular carcinoma in CuZn superoxide dismutase deficient mice PROTEOMICS Elchuri, S., Naeemuddin, M., Sharpe, O., Robinson, W. H., Huang, T. 2007; 7 (12): 2121-2129

  Abstract

  To identify biomarkers associated with the development of hepatocellular carcinoma (HCC) in CuZn superoxide dismutase (CuZnSOD, Sod1) deficient mice, 2-DE followed by MS analysis was carried out with liver samples obtained from 18-month-old Sod1-/- and +/+ mice. The intracellular Ca binding protein, regucalcin (RGN), showed a divergent alteration in Sod1-/- samples. Whereas elevated RGN levels were observed in -/- samples with no obvious neoplastic changes, marked reduction in RGN was observed in -/- samples with fully developed HCC. GST mu1 (GSTM1), on the other hand, showed a significant increase only in the neoplastic regions obtained from Sod1-/- livers. No change in GSTM1 was observed in the surrounding normal tissues. Marked reduction was observed in two intracellular lipid transporters, fatty acid binding protein 1 (FABP1) and major urinary protein 11 and 8 (MUP 11&8), in Sod1-/- samples. Analysis of additional samples at 18-22 months of age showed a three-fold increase in enolase activities in Sod1-/- livers. Consistent with previous findings, carbonic anhydrase 3 (CAIII) levels were significantly reduced in Sod1-/- samples, and immunohistochemical analysis revealed that the reduction was not homogenous throughout the lobular structure in the liver.

  View details for DOI 10.1002/pmic.200601011

  View details for PubMedID 17514684

• Imatinib for the treatment of rheumatic diseases NATURE CLINICAL PRACTICE RHEUMATOLOGY Paniagua, R. T., Robinson, W. H. 2007; 3 (4): 190-191

  View details for DOI 10.1038/ncprheum0465

  View details for PubMedID 17396105

• Suppressive Role of CD72 in Experimental Autoimmune Encephalomyelitis Li, D. H., Creusot, R., Ousman, S. S., Fontoura, P., Sobel, R. A., Cao, T. M., Ho, P., Youssef, S., Robinson, W. H., Hong, A., Steinman, L., Parnes, J. R. AMER ASSOC IMMUNOLOGISTS. 2007

  View details for Web of Science ID 000209758203196

• Signal transduction blockade with imatinib mesylate prevents and treats autoimmune arthritis Paniagua, R., Sharpe, O., Ho, P., Chan, S., Chang, A., Tomooka, B., Lee, D., Genovese, M., Utz, P., Steinman, L., Robinson, W. AMER ASSOC IMMUNOLOGISTS. 2007

  View details for Web of Science ID 000209758200096

• Protective and therapeutic role for alpha B-crystallin in autoimmune demyelination Ousman, S., Tomooka, B., Van Noort, J., O'Conner, K., Wawrousek, E., Hafler, D., Sobel, R., Robinson, W., Steinman, L. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2007: S139

  View details for DOI 10.1016/j.clim.2007.03.034

  View details for Web of Science ID 000247137200362

• Proteomic signatures of secreted proteins for the prediction of treatment response to TNF blockers in RA Hueber, W., Tomooka, B., Batliwalla, F., Tibshirani, R., Gregersen, P., Klareskog, L., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2007: S94

  View details for DOI 10.1016/j.clim.2007.03.449

  View details for Web of Science ID 000247137200245

• Tolerizing DNA vaccines for autoimmune arthritis AUTOIMMUNITY Ho, P. P., Higgins, J. P., Kidd, B. A., Tomooka, B., Digennaro, C., Lee, L. Y., De Vegvar, H. E., Steinman, L., Robinson, W. H. 2006; 39 (8): 675-682

  Abstract

  Current therapies for rheumatoid arthritis (RA) and other autoimmune diseases non-specifically suppress immune function, and there is great need for fundamental approaches such as antigen-specific tolerizing therapy. In this paper we describe development of antigen-specific tolerizing DNA vaccines to treat collagen-induced arthritis (CIA) in mice, and use of protein microarrays to monitor response to therapy and to identify potential additional autoimmune targets for next generation vaccines. We demonstrate that tolerizing DNA vaccines encoding type II collagen (CII) reduced the incidence and severity of CIA. Atorvastatin, a statin drug found to reduce the severity of autoimmunity, potentiated the effect of DNA vaccines encoding CII. Analysis of cytokines produced by collagen-reactive T cells derived from mice receiving tolerizing DNA encoding CII, as compared to control vaccines, revealed reduced production of the pro-inflammatory cytokines IFN-gamma and TNF-alpha. Arthritis microarray analysis demonstrated reduced spreading of autoantibody responses in mice treated with DNA encoding CII. The development of tolerizing DNA vaccines, and the use of antibody profiling to guide design of and to monitor therapeutic responses to such vaccines, represents a promising approach for the treatment of RA and other autoimmune diseases.

  View details for DOI 10.1016/08916930601061603

  View details for PubMedID 17178564

• Synovial proteome microarray identifies citrullinated vimentin as an autoantigen in DR4-IE tg mice following immunization with citrullinated human fibrinogen. Brintnell, W., Hill, J. A., Bell, D. A., Hueber, W., Tomooka, B. H., Robinson, W. H., Cairns, E. WILEY-LISS. 2006: S179

  View details for Web of Science ID 000240877200388

• Monoclonal antibodies to myelin proteolipid protein (PLP) epitopes inhibit neurite outgrowth: Implications for MS Hinojoza, J. R., Han, M. H., Eaton, M., Sharpe, O., Steinman, L., Robinson, W. H., Sobel, R. A. BLACKWELL PUBLISHING. 2006: S13

  View details for Web of Science ID 000239938600027

• Specific cytokine and chemokine increases precede the appearance of anti-cyclic citrullinated protein (Anti-CCP) antibodies and rheumatoid factor (RF) in the pre-clinical period of rheumatoid arthritis (RA) development. Deane, K. D., Hueber, W., Majka, D. S., Tomooka, B. H., Gilliland, W. R., Parrish, L. A., Parish, M. C., Norris, J. M., Robinson, W. H., Holers, V. WILEY-LISS. 2006: S508

  View details for Web of Science ID 000240877202356

• Proteomic biomarkers for autoimmune disease PROTEOMICS Hueber, W., Robinson, W. H. 2006; 6 (14): 4100-4105

  Abstract

  Autoimmune diseases affect 3% of the world population, yet the diagnosis and classification of autoimmune diseases remain based on clinical examination combined with traditional laboratory tests and imaging studies. The development of genomic and proteomic technologies provides an unprecedented ability to identify novel biosignatures to diagnose, classify, and guide therapeutic decision making in patients with autoimmune disease. In this article, we review recent advances in proteomics technologies and their application to autoimmune disease.

  View details for DOI 10.1002/pmic.200600017

  View details for PubMedID 16786488

• Fluid supported lipid bilayers containing mono sialoganglioside GM1: A QCM-D and FRAP study COLLOIDS AND SURFACES B-BIOINTERFACES Weng, K. C., Kanter, J. L., Robinson, W. H., Frank, C. W. 2006; 50 (1): 76-84

  Abstract

  In an effort to use model fluid membranes for immunological studies, we compared the formation of planar phospholipid bilayers supported on silicon dioxide surfaces with and without incorporation of glycolipids as the antigen for in situ antibody binding. Dynamic light scattering measurements did not differentiate the hydrodynamic volumes of extruded small unilamellar vesicles (E-SUVs) containing physiologically relevant concentrations (0.5-5 mol%) of monosialoganglioside GM1 (GM1) from exclusive egg yolk L-alpha-phosphatidylcholine (egg PC) E-SUVs. However, quantifiable differences in deposition mass and dissipative energy loss emerged in the transformation of 5 mol% GM1/95 mol% egg PC E-SUVs to planar supported lipid bilayers (PSLBs) by vesicle fusion on thermally evaporated SiO2, as monitored by the quartz crystal microbalance with dissipation (QCM-D) technique. Compared to the 100 mol% egg PC bilayers on the same surface, E-SUVs containing 5 mol% GM1 reached a approximately 12% higher mass and a lower dissipative energy loss during bilayer transformation. PSLBs with 5 mol% GM1 are approximately 18% heavier than 100 mol% egg PC and approximately 11% smaller in projected area per lipid, indicating an increased rigidity and a tighter packing. Subsequent binding of polyclonal immunoglobulin G anti-GM1 to the PSLBs was performed in situ and showed specificity. The anti-GM1 to GM1 ratios at equilibrium were roughly proportional to the concentrations of anti-GM1 administered in the solution. Fluorescence recovery after photobleaching was utilized to verify the retained, albeit reduced lateral fluidity of the supported membranes. Five moles percentage of GM1 membranes (GM1 to PC ratio approximately 1:19) decorated with 1 mol% N-(Texas Red sulfonyl)-1,2-dihexadecanoyl-sn-glycerol-3-phosphoethanolamine (Texas Red DHPE) exhibited an approximately 16% lower diffusion coefficient of 1.32+/-0.06 microm2/s, compared to 1.58+/-0.04 microm2/s for egg PC membranes without GM1 (p<0.01). The changes in vesicle properties and membrane lateral fluidity are attributed to the interactions of GM1 with itself and GM1 with other membrane lipids. This system allows for molecules of interest such as GM1 to exist on a more biologically relevant surface than those used in conventional methods such as ELISA. Our analysis of rabbit serum antibodies binding to GM1 demonstrates this platform can be used to test for the presence of anti-lipid antibodies in serum.

  View details for DOI 10.1016/j.colsurfb.2006.03.010

  View details for PubMedID 16730958

• Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis JOURNAL OF CLINICAL INVESTIGATION Kuhn, K. A., Kulik, L., Tomooka, B., Braschler, K. J., Arend, W. P., Robinson, W. H., Holers, V. M. 2006; 116 (4): 961-973

  Abstract

  Antibodies against citrullinated proteins are specific and predictive markers for rheumatoid arthritis although the pathologic relevance of these antibodies remains unclear. To investigate the significance of these autoantibodies, collagen-induced arthritis (CIA) in mice was used to establish an animal model of antibody reactivity to citrullinated proteins. DBA/1J mice were immunized with bovine type II collagen (CII) at days 0 and 21, and serum was collected every 7 days for analysis. Antibodies against both CII and cyclic citrullinated peptide, one such citrullinated antigen, appeared early after immunization, before joint swelling was observed. Further, these antibodies demonstrated specific binding to citrullinated filaggrin in rat esophagus by indirect immunofluorescence and citrullinated fibrinogen by Western blot. To evaluate the role of immune responses to citrullinated proteins in CIA, mice were tolerized with a citrulline-containing peptide, followed by antigen challenge with CII. Tolerized mice demonstrated significantly reduced disease severity and incidence compared with controls. We also identified novel murine monoclonal antibodies specific to citrullinated fibrinogen that enhanced arthritis when coadministered with a submaximal dose of anti-CII antibodies and bound targets within the inflamed synovium of mice with CIA. These results demonstrate that antibodies against citrullinated proteins are centrally involved in the pathogenesis of autoimmune arthritis.

  View details for DOI 10.1172/JCI25422

  View details for Web of Science ID 000236556100018

  View details for PubMedID 16585962

  View details for PubMedCentralID PMC1421345

• Lipid microarrays identify key mediators of autoimmune brain inflammation Kanter, J., Johnson, A., Ho, P. P., Hur, E., Narayana, S., Robinson, W., Steinman, L. AMER ASSOC IMMUNOLOGISTS. 2006: S145

  View details for Web of Science ID 000238837101165

• Antigen arrays for antibody profiling CURRENT OPINION IN CHEMICAL BIOLOGY Robinson, W. H. 2006; 10 (1): 67-72

  Abstract

  Antigen array technologies enable large-scale profiling of the specificity of antibody responses against autoantigens, tumor antigens and microbial antigens. Antibody profiling will provide insights into pathogenesis, and will enable development of novel tests for diagnosis and guiding therapy in the clinic. Recent advances in the field include development of antigen array-based approaches to examine immune responses against antigens encoded in genetic libraries, post-translationally modified proteins, and other biomolecules such as lipids. A promising application is the use of antibody profiling to guide development and selection of antigen-specific therapies to treat autoimmune disease. This review discusses these advances and the challenges ahead for development and refinement of antibody profiling technologies for use in the research laboratory and the clinic.

  View details for DOI 10.1016/j.cbpa.2005.12.028

  View details for PubMedID 16406767

• Microarray identified lipids modulate autoimmune demyelinating disease. Kanter, J., Johnson, A., Ho, P., Hur, E., Narayana, S., Steinman, L., Robinson, W. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2006: S29

  View details for DOI 10.1016/j.clim.2006.04.223

  View details for Web of Science ID 000237924300068

• Fibrinogen-induced arthritis in mice as a model for rheumatoid arthritis. Robinson, W., Kuhn, K., Tomooka, B., Holers, V., Ho, P. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2006: S16–S17

  View details for DOI 10.1016/j.clim.2006.04.037

  View details for Web of Science ID 000237924300036

• Treatment of autoimmune neuroinflammation with a synthetic tryptophan metabolite SCIENCE Platten, M., Ho, P. P., Youssef, S., Fontoura, P., Garren, H., Hur, E. M., Gupta, R., Lee, L. Y., Kidd, B. A., Robinson, W. H., Sobel, R. A., Selley, M. L., Steinman, L. 2005; 310 (5749): 850-855

  Abstract

  Local catabolism of the amino acid tryptophan (Trp) by indoleamine 2,3-dioxygenase (IDO) is considered an important mechanism of regulating T cell immunity. We show that IDO transcription was increased when myelin-specific T cells were stimulated with tolerogenic altered self-peptides. Catabolites of Trp suppressed proliferation of myelin-specific T cells and inhibited production of proinflammatory T helper-1 (T(H)1) cytokines. N-(3,4,-Dimethoxycinnamoyl) anthranilic acid (3,4-DAA), an orally active synthetic derivative of the Trp metabolite anthranilic acid, reversed paralysis in mice with experimental autoimmune encephalomyelitis, a model of multiple sclerosis (MS). Trp catabolites and their derivatives offer a new strategy for treating T(H)1-mediated autoimmune diseases such as MS.

  View details for DOI 10.1126/science.1117634

  View details for PubMedID 16272121

• A suppressive oligodeoxynucleotide enhances the efficacy of myelin cocktail/IL-4-tolerizing DNA vaccination and treats autoimmune disease JOURNAL OF IMMUNOLOGY Ho, P. P., Fontoura, P., Platten, M., Sobel, R. A., DeVoss, J. J., Lee, L. Y., Kidd, B. A., Tomooka, B. H., Capers, J., Agrawal, A., Gupta, R., Zernik, J., Yee, M. K., Lee, B. J., Garren, H., Robinson, W. H., Steinman, L. 2005; 175 (9): 6226-6234

  Abstract

  Targeting pathogenic T cells with Ag-specific tolerizing DNA vaccines encoding autoantigens is a powerful and feasible therapeutic strategy for Th1-mediated autoimmune diseases. However, plasmid DNA contains abundant unmethylated CpG motifs, which induce a strong Th1 immune response. We describe here a novel approach to counteract this undesired side effect of plasmid DNA used for vaccination in Th1-mediated autoimmune diseases. In chronic relapsing experimental autoimmune encephalomyelitis (EAE), combining a myelin cocktail plus IL-4-tolerizing DNA vaccine with a suppressive GpG oligodeoxynucleotide (GpG-ODN) induced a shift of the autoreactive T cell response toward a protective Th2 cytokine pattern. Myelin microarrays demonstrate that tolerizing DNA vaccination plus GpG-ODN further decreased anti-myelin autoantibody epitope spreading and shifted the autoreactive B cell response to a protective IgG1 isotype. Moreover, the addition of GpG-ODN to tolerizing DNA vaccination therapy effectively reduced overall mean disease severity in both the chronic relapsing EAE and chronic progressive EAE mouse models. In conclusion, suppressive GpG-ODN effectively counteracted the undesired CpG-induced inflammatory effect of a tolerizing DNA vaccine in a Th1-mediated autoimmune disease by skewing both the autoaggressive T cell and B cell responses toward a protective Th2 phenotype. These results demonstrate that suppressive GpG-ODN is a simple and highly effective novel therapeutic adjuvant that will boost the efficacy of Ag-specific tolerizing DNA vaccines used for treating Th1-mediated autoimmune diseases.

  View details for PubMedID 16237121

• Antigen microarray profiling of autoantibodies in rheumatoid arthritis ARTHRITIS AND RHEUMATISM Hueber, W., Kidd, B. A., Tomooka, B. H., Lee, B. J., Bruce, B., Fries, J. F., Sonderstrup, G., Monach, P., Drijfhout, J. W., van Venrooij, W. J., Utz, P. J., Genovese, M. C., Robinson, W. H. 2005; 52 (9): 2645-2655

  Abstract

  Because rheumatoid arthritis (RA) is a heterogeneous autoimmune disease in terms of disease manifestations, clinical outcomes, and therapeutic responses, we developed and applied a novel antigen microarray technology to identify distinct serum antibody profiles in patients with RA.Synovial proteome microarrays, containing 225 peptides and proteins that represent candidate and control antigens, were developed. These arrays were used to profile autoantibodies in randomly selected sera from 2 different cohorts of patients: the Stanford Arthritis Center inception cohort, comprising 18 patients with established RA and 38 controls, and the Arthritis, Rheumatism, and Aging Medical Information System cohort, comprising 58 patients with a clinical diagnosis of RA of <6 months duration. Data were analyzed using the significance analysis of microarrays algorithm, the prediction analysis of microarrays algorithm, and Cluster software.Antigen microarrays demonstrated that autoreactive B cell responses targeting citrullinated epitopes were present in a subset of patients with early RA with features predictive of the development of severe RA. In contrast, autoimmune targeting of the native epitopes contained on synovial arrays, including several human cartilage gp39 peptides and type II collagen, were associated with features predictive of less severe RA.Proteomic analysis of autoantibody reactivities provides diagnostic information and allows stratification of patients with early RA into clinically relevant disease subsets.

  View details for DOI 10.1002/art.21269

  View details for PubMedID 16142722

• Fibrinogen-induced arthritis in mice as a novel model for rheumatoid arthritis. Robinson, W. H., Tomooka, B., Genovese, M. C., Ho, P. P. WILEY-BLACKWELL. 2005: S157

  View details for Web of Science ID 000232207800353

• Integrated serum proteome analysis of autoantibodies and cytokines delineates distinct biosignatures in subsets of patients with early RA. Hueber, W., Tomooka, B., Bruce, B., Monach, P., Fries, J. F., van Venrooij, W., Zatarain, E., Genovese, M. C., Robinson, W. H. WILEY-BLACKWELL. 2005: S276–S277

  View details for Web of Science ID 000232207801206

• Lipid microarrays detect diverse anti-lipid antibodies in MS and EAE: Sulfatide immunization exacerbates EAE Kanter, J. L., Narayana, S., Ho, P., Sobel, R., Steinman, L., Robinson, W. H. FEDERATION AMER SOC EXP BIOL. 2005: A1441

  View details for Web of Science ID 000227610903189

• Suppression of autoimmunity via microbial mimics of altered peptide ligands MOLECULAR MIMICRY: INFECTION-INDUCING AUTOIMMUNE DISEASE Steinman, L., Utz, P. J., Robinson, W. H. 2005; 296: 55-63

  Abstract

  Molecular mimics of self-antigens can behave as altered peptide ligands and serve to ameliorate autoimmune disease. Analysis of experimental autoimmune encephalomyelitis with proteomic autoantibody microarrays reveals that there might exist a wide variety of microbes with features that mimic self-epitopes. Autoimmunity could therefore be modulated via microbial immunity, which may account for relapse and remission of ongoing disease.

  View details for Web of Science ID 000235217000004

  View details for PubMedID 16323420

• Immunity to the extracellular domain of Nogo-A modulates experimental autoimmune encephalomyelitis JOURNAL OF IMMUNOLOGY Fontoura, P., Ho, P. P., DeVoss, J., Zheng, B. H., Lee, B. J., Kidd, B. A., Garren, H., Sobel, R. A., Robinson, W. H., Tessier-Lavigne, M., Steinman, L. 2004; 173 (11): 6981-6992

  Abstract

  Nogo-66, the extracellular 66 aa loop of the Nogo-A protein found in CNS myelin, interacts with the Nogo receptor and has been proposed to mediate inhibition of axonal regrowth. It has been shown that immunization with Nogo-A promotes recovery in animal models of spinal cord injury through induction of Ab production. In this report, studies were performed to characterize the immune response to Nogo-66 and to determine the role of Nogo in experimental autoimmune encephalomyelitis (EAE). Immunization of EAE-susceptible mouse strains with peptides derived from Nogo-66 induced a CNS immune response with clinical and pathological similarities to EAE. The Nogo-66 peptides elicited strong T cell responses that were not cross-reactive to other encephalitogenic myelin Ags. Using a large scale spotted microarray containing proteins and peptides derived from a wide spectrum of myelin components, we demonstrated that Nogo-66 peptides also generated a specific Ab response that spreads to several other encephalitogenic myelin Ags following immunization. Nogo-66-specific T cell lines ameliorated established EAE, via Nogo-66-specific Th2 cells that entered the CNS. These results indicate that some T cell and B cell immune responses to Nogo-66 are associated with suppression of ongoing EAE, whereas other Nogo-66 epitopes can be encephalitogenic.

  View details for PubMedID 15557195

• Characterization of novel antigens recognized by serum autoantibodies from anti-CD1 TCR-transgenic lupus mice EUROPEAN JOURNAL OF IMMUNOLOGY Hueber, W., Zeng, D. F., Sharpe, O., Robinson, W. H., Strober, S., Utz, P. J. 2004; 34 (6): 1654-1662

  Abstract

  In this study, we further characterize the humoral autoimmune response in the recently described anti-CD1 autoreactive T cell receptor-transgenic mouse lupus model (CD1 lupus model). We discovered and characterized novel autoantigens, comprising a protein of 105 kDa (p105) and a novel RNA molecule of 140 base pairs (bp) that is likely associated with p105, and several additional factors with distinct biochemical properties. In the CD1 lupus model, lethally irradiated BALB/c/nu/nu mice were injected intravenously with sorted bone marrow cells and sorted splenic T cells from donor BALB/c mice expressing TCR alpha and beta transgenes that encode autoreactivity for CD1d. Adoptive hosts injected with the single-positive (CD4(+) and CD8(+)) subset of transgenic cells developed anti-double-stranded DNA antibodies and a lupus-like illness. Sera were analyzed by Western blotting and immunoprecipitation. Antigens were characterized by biochemical and serological methods. Serum autoantibodies from 5 of 12 (42%) CD1 lupus mice immunoprecipitated a 105-kDa protein, termed p105. p105 was associated with a small RNA of approximately 140 bp. Anti-p105 autoantibodies appeared early in the course of disease. Serological and biochemical characterization suggested that p105 was distinct from known lupus autoantigens of similar molecular masses, indicating that p105 represents a novel autoantigen in lupus.

  View details for DOI 10.1002/eji.200324201

  View details for PubMedID 15162435

• High-throughput methods for measuring autoantibodies in systemic lupus erythematosus and other autoimmune diseases AUTOIMMUNITY Graham, K. L., Robinson, W. H., Steinman, L., Utz, P. J. 2004; 37 (4): 269-272

  Abstract

  Numerous groups have now validated high-throughput approaches to autoantibody profiling in a variety of systems. Recently, we have used autoantigen microarray technology to identify distinct autoantibody profiles in H-2 congenic MRL/lpr mice (Sekine et al., manuscript in preparation), and we are expanding this platform to study human and mouse models of IDDM and RA. We are also developing protein arrays for multiplex analysis of serum antibody isotypes. Multiplexed methods for autoantibody profiling will undoubtedly continue to uncover novel aspects of autoimmunity and B cell biology. It is now time to move these technologies beyond the proof-of-concept phase, and start addressing the next series of important questions. These include, but certainly are not limited to: identifying "autoantibody signatures" associated with disease state or outcome; profiling autoantibodies during the natural course of murine and human disease; and monitoring changes in autoantibody profiles of patients in response to therapeutic intervention. However, the next set of challenges is just right around the corner. As data and statistical analysis tools become more robust, it will be possible to generate and approach new hypotheses at an unprecedented pace.

  View details for DOI 10.1080/08916930410001710686

  View details for Web of Science ID 000223372000006

  View details for PubMedID 15518040

• Microarray profiling of antiviral antibodies for the development of diagnostics, vaccines, and therapeutics CLINICAL IMMUNOLOGY de Vegvar, H. E., Robinson, W. H. 2004; 111 (2): 196-201

  Abstract

  Multiplex analysis of antiviral antibody (Ab) responses provides a potentially powerful strategy for viral diagnosis, prognostication, and development of vaccines and prophylactic Abs. In the coming years, advancements in proteomic technologies will provide even more robust methods to characterize antiviral Ab responses. Biomedical researchers will be faced with the exciting challenge of identifying antiviral Ab specificities that correlate with improved outcomes and efficacious interventions, and translating the findings into more effective diagnostics, prophylactics, and therapeutics.

  View details for PubMedID 15137952

• Unlocking the "PAD" lock on rheumatoid arthritis ANNALS OF THE RHEUMATIC DISEASES Utz, P. J., Genovese, M. C., Robinson, W. H. 2004; 63 (4): 330-332

  View details for DOI 10.1136/ard.2003.015990

  View details for Web of Science ID 000220198000002

  View details for PubMedID 15020322

  View details for PubMedCentralID PMC1754966

• Genomic and proteomic analysis of multiple sclerosis - Opinion CURRENT OPINION IN IMMUNOLOGY Robinson, W. H., Utz, P. J., Steinman, L. 2003; 15 (6): 660-667

  Abstract

  Multiple sclerosis (MS) and other autoimmune diseases result from the dysregulation of genetic and proteomic programs. In MS, the loss of immune homeostasis leads to aberrant targeting and destruction of the myelin sheath, which manifests as the clinical syndrome of MS. The advent of technologies to perform large-scale analysis of mRNA transcript and protein expression will transform our understanding of the mechanisms underlying the initiation and progression of MS, and will yield new targets for therapeutic intervention.

  View details for DOI 10.1016/j.coi.2003.09.015

  View details for PubMedID 14630200

• Protein arrays for autoantibody profiling and fine-specificity mapping PROTEOMICS Robinson, W. H., Steinman, L., Utz, P. J. 2003; 3 (11): 2077-2084

  Abstract

  Protein arrays provide a powerful approach to study autoimmune disease. Autoimmune responses activate B cells to produce autoantibodies that recognize self-molecules termed autoantigens, many of which are proteins or protein complexes. Protein arrays enable profiling of the specificity of autoantibody responses against panels of peptides and proteins representing known autoantigens as well as candidate autoantigens. In addition to identifying autoantigens and mapping immunodominant epitopes, proteomic analysis of autoantibody responses will further enable diagnosis, prognosis, and tailoring of antigen-specific tolerizing therapy.

  View details for DOI 10.1002/pmic.200300583

  View details for PubMedID 14595805

• Microarray profiling of antibody responses against simian-human immunodeficiency virus: Postchallenge convergence of reactivities independent of host histocompatibility type and vaccine regimen JOURNAL OF VIROLOGY de Vegvar, H. E., Amara, R. R., Steinman, L., Utz, P. J., Robinson, H. L., Robinson, W. H. 2003; 77 (20): 11125-11138

  Abstract

  We developed antigen microarrays to profile the breadth, strength, and kinetics of epitope-specific antiviral antibody responses in vaccine trials with a simian-human immunodeficiency virus (SHIV) model for human immunodeficiency virus (HIV) infection. These arrays contained 430 distinct proteins and overlapping peptides spanning the SHIV proteome. In macaques vaccinated with three different DNA and/or recombinant modified vaccinia virus Ankara (rMVA) vaccines encoding Gag-Pol or Gag-Pol-Env, these arrays distinguished vaccinated from challenged macaques, identified three novel viral epitopes, and predicted survival. Following viral challenge, anti-SHIV antibody responses ultimately converged to target eight immunodominant B-cell regions in Env regardless of vaccine regimen, host histocompatibility type, and divergent T-cell specificities. After challenge, responses to nonimmunodominant epitopes were transient, while responses to dominant epitopes were gained. These data suggest that the functional diversity of anti-SHIV B-cell responses is highly limited in the presence of persisting antigen.

  View details for DOI 10.1128/JVI.77.20.11125-11138.2003

  View details for PubMedID 14512560

• Autoantibodies in early arthritis: Advances in diagnosis and prognostication CLINICAL AND EXPERIMENTAL RHEUMATOLOGY Hueber, W., Utz, P. J., Robinson, W. H. 2003; 21 (5): S59-S64

  Abstract

  Several excellent reviews have recently been published on the significance of autoantibodies in rheumatoid arthritis (RA) (1-4). Here we: (i) review selected longitudinal studies examining the predictive utility of autoantibodies in early arthritis and early RA cohorts; (ii) assess the relevance of autoantibodies as an independent parameter for prediction and prognostication of RA; and (iii) describe the potential of multiplex autoantibody assays, including miniaturized, high-throughput microarray technology, to improve diagnosis and prognostication in recent-onset synovitis/early arthritis patients.

  View details for Web of Science ID 000185970100011

  View details for PubMedID 14969052

• Protein and peptide array analysis of autoimmune disease BIOTECHNIQUES Robinson, W. H., Steinman, L., Utz, P. J. 2002: 66-69

  Abstract

  Molecular cloning, sequencing of the human genome, and other major advances in biomedical research have contributed substantially to our understanding of autoimmune disease. Nevertheless, to date, such advances have failed to reveal the etiology of or yield curative therapies for autoimmune disease. New approaches are needed. Proteomics, the large-scale study of expression and function of proteins that compose our tissues and mediate disease, represents a powerful and promising strategy. We developed protein and peptide arrays to profile autoantibody responses in autoimmune disease. Protein and peptide array analysis of autoimmune samples is revealing human and pathogen proteins involved in initiation and perpetuation of autoimmunity. Proteomic determination of autoantibody profiles can be utilized for diagnosis, prognostication, and guiding tolerizing therapy for autoimmune disease.

  View details for PubMedID 12514932

• Millennium Award. Proteomics for the development of DNA tolerizing vaccines to treat autoimmune disease. Clinical immunology Robinson, W. H., Garren, H., Utz, P. J., Steinman, L. 2002; 103 (1): 7-12

  Abstract

  Autoimmune disease affects 3% of the world population, yet current therapies that globally suppress immune function are inadequate. Tremendous need exists for specific and curative therapies, and we describe a strategy for development of antigen-specific therapies that inactivate pathogenic lymphocytes causing tissue injury. Major barriers to development of antigen-specific therapies for T-cell-mediated autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis, and autoimmune diabetes, include (i) lack of knowledge of the specificity of autoimmune responses, for which proteomic technologies represent powerful tools to identify the self-protein targets of the autoimmune response, and (ii) lack of methods to induce specific immune tolerance, for which DNA tolerizing vaccines represent a promising strategy. We termed our approach Reverse Genomics: use of the proteomics-determined specificity of the autoantibody response to develop and select DNA tolerizing vaccines. Studies performed using animal models for multiple sclerosis and autoimmune diabetes support our Reverse Genomics approach. Through integration of proteomics with specific tolerizing therapies, we are developing a comprehensive approach to treat human autoimmune disease.

  View details for PubMedID 11987980

• Proteomics for the development of DNA tolerizing vaccines to treat autoimmune disease CLINICAL IMMUNOLOGY Robinson, W. H., Garren, H., Utz, P. J., Steinman, L. 2002; 103 (1): 7-12

  Abstract

  Autoimmune disease affects 3% of the world population, yet current therapies that globally suppress immune function are inadequate. Tremendous need exists for specific and curative therapies, and we describe a strategy for development of antigen-specific therapies that inactivate pathogenic lymphocytes causing tissue injury. Major barriers to development of antigen-specific therapies for T-cell-mediated autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis, and autoimmune diabetes, include (i) lack of knowledge of the specificity of autoimmune responses, for which proteomic technologies represent powerful tools to identify the self-protein targets of the autoimmune response, and (ii) lack of methods to induce specific immune tolerance, for which DNA tolerizing vaccines represent a promising strategy. We termed our approach Reverse Genomics: use of the proteomics-determined specificity of the autoantibody response to develop and select DNA tolerizing vaccines. Studies performed using animal models for multiple sclerosis and autoimmune diabetes support our Reverse Genomics approach. Through integration of proteomics with specific tolerizing therapies, we are developing a comprehensive approach to treat human autoimmune disease.

  View details for DOI 10.1006/clim.2002.5185

  View details for Web of Science ID 000175409000002

• Proteomics technologies for the study of autoimmune disease ARTHRITIS AND RHEUMATISM Robinson, W. H., Steinman, L., Utz, P. J. 2002; 46 (4): 885-893

  View details for PubMedID 11953963

• Autoantibody profiling for the study and treatment of autoimmune disease ARTHRITIS RESEARCH Hueber, W., Utz, P. J., Steinman, L., Robinson, W. H. 2002; 4 (5): 290-295

  Abstract

  Proteomics technologies enable profiling of autoantibody responses using biological fluids derived from patients with autoimmune disease. They provide a powerful tool to characterize autoreactive B-cell responses in diseases including rheumatoid arthritis, multiple sclerosis, autoimmune diabetes, and systemic lupus erythematosus. Autoantibody profiling may serve purposes including classification of individual patients and subsets of patients based on their 'autoantibody fingerprint', examination of epitope spreading and antibody isotype usage, discovery and characterization of candidate autoantigens, and tailoring antigen-specific therapy. In the coming decades, proteomics technologies will broaden our understanding of the underlying mechanisms of and will further our ability to diagnose, prognosticate and treat autoimmune disease.

  View details for PubMedID 12223102

• Demyelinating and neurologic events reported in association with tumor necrosis factor alpha antagonism - By what mechanisms could tumor necrosis factor alpha antagonists improve rheumatoid arthritis but exacerbate multiple sclerosis? ARTHRITIS AND RHEUMATISM Robinson, W. H., Genovese, M. C., Moreland, L. W. 2001; 44 (9): 1977-1983

  View details for Web of Science ID 000172491200004

  View details for PubMedID 11592357

• Protein array characterization of the specificity of the autoantibody response in experimental autoimmune encephalomyelitis: Examination of the role of epitope spreading in disease progression Robinson, W. H., DiGennaro, C., Garren, H., Hirschberg, D., Haab, B., Brown, P., Utz, P. J., Steinman, L. FEDERATION AMER SOC EXP BIOL. 2001: A1064

  View details for Web of Science ID 000167454202030

• Allele-specific expression of the mouse B-cell surface protein CD72 on T cells IMMUNOGENETICS Robinson, W. H., LANDOLFI, M. M., Parnes, J. R. 1997; 45 (3): 195-200

  Abstract

  CD72 is a 45 000 Mr mouse B-cell surface glycoprotein involved in B-cell proliferation and differentiation. Expression of mouse CD72 is thought to be restricted to the B-cell lineage. We recently demonstrated that the monoclonal antibodies K10.6 and B9.689, previously defined as recognizing the mouse lymphocyte alloantigens Ly-19.2 and Ly-32.2, respectively, recognize specific alleles of CD72. Early studies using antibody-mediated cytotoxicity assays demonstrated that K10.6 and B9.689 react with B cells, several T-cell lines, and a subset of peripheral T cells. These findings led us to consider the possibility that CD72 might also be expressed on a subset of T cells. In this report we demonstrate that CD72 is constitutively expressed on a fraction of peripheral T cells isolated from strains of mice expressing the CD72(b) allele, but not the CD72(a) or CD72(c) alleles. Three days after activating T cells with concanavalin A or plate-bound CD3-specific mAb, CD72 is expressed on a larger fraction of peripheral T cells as well as a fraction of thymocytes from mouse strains expressing the CD72(b) allele. CD72 is expressed on both the CD4(+) and CD8(+) thymocyte and peripheral T-cell subsets. No CD72 expression is detected on activated thymocytes or peripheral T cells from mouse strains expressing the CD72(a) or CD72(c) alleles. Expression of CD72(b) on peripheral T cells was confirmed by northern blot analysis demonstrating CD72 mRNA expression. These results demonstrate that CD72 expression is not restricted to B lineage cells in mouse strains expressing the CD72(b) allele; instead, a population of T lineage cells in these mice also expresses CD72.

  View details for Web of Science ID A1997WE19000005

  View details for PubMedID 8995186

• IDENTIFICATION OF A MOUSE PROTEIN HOMOLOGOUS TO THE HUMAN CD6 T-CELL SURFACE PROTEIN AND SEQUENCE OF THE CORRESPONDING CDNA JOURNAL OF IMMUNOLOGY Robinson, W. H., Prohaska, S. S., Santoro, J. C., Robinson, H. L., Parnes, J. R. 1995; 155 (10): 4739-4748

  Abstract

  CD6 is a 105/130 kDa monomeric T cell surface glycoprotein that has been shown to play a role in human T cell activation. Recently a partial mouse CD6 cDNA sequence was described. We have isolated full-length cDNA clones including the initiation codon and sequence encoding the full signal peptide, as well as an additional 39 amino acids within the cytoplasmic domain as compared to the previously reported clone. The predicted full-length mouse CD6 protein contains 665 amino acids and has the features of a type I integral membrane protein. The extracellular domain of mouse CD6 is composed of three repeated cysteine-rich domains similar to those in human CD6, mouse and human CD5, and other members of a family of proteins whose prototype is the type I macrophage scavenger receptor. In marked contrast to the previously published human CD6 sequence, the mouse sequence predicts a long cytoplasmic tail that is not closely related to other proteins and possesses two proline-rich motifs containing the SH3-domain binding consensus sequence, three protein kinase C phosphorylation site motifs, nine casein kinase-2 phosphorylation site motifs, and a serine-threonine-rich motif repeated three times. Northern blot analysis revealed that mouse CD6 mRNA is expressed predominantly in thymus, lymph node, and spleen. A polyclonal antiserum was raised against mouse CD6 by gene gun plasmid DNA immunization of rabbits with the mouse CD6 cDNA in an expression vector. In immunofluorescence analysis this polyclonal antiserum positively stained the surface of cells transfected with the mouse CD6 cDNA in an expression vector, as well as most normal mouse thymocytes and peripheral T cells. CD6 protein is expressed on most CD4+CD8+ double-positive and CD4+ or CD8+ single-positive thymocytes, and is expressed at highest levels on mature CD3high thymocytes. The expression of mouse CD6 in thymocytes and peripheral T cells correlates closely with the expression of the related CD5 molecule. The polyclonal rabbit anti-mouse CD6 Abs immunoprecipitated a major polypeptide of 128 kDa from resting and 130 kDa from PMA- and FCS-activated mouse thymocytes and lymph node cells; it is likely that this increase in size upon activation is due to phosphorylation of mouse CD6 as has been described for human CD6. These data demonstrate that mouse thymocytes and T cells express a 130-kDa cell surface protein homologous to human CD6.

  View details for Web of Science ID A1995TD88800029

  View details for PubMedID 7594475

• HUMAN CD6 POSSESSES A LARGE, ALTERNATIVELY SPLICED CYTOPLASMIC DOMAIN EUROPEAN JOURNAL OF IMMUNOLOGY Robinson, W. H., DEVEGVAR, H. E., Prohaska, S. S., Rhee, J. W., Parnes, J. R. 1995; 25 (10): 2765-2769

  Abstract

  Human CD6 is a monomeric 105/130-kDa T cell surface glycoprotein that is involved in T cell activation. The apparent discrepancy between the size of the cytoplasmic domain in human (44 amino acids) and mouse (243 amino acids) CD6, led us to use reverse transcriptase-polymerase chain reaction of human peripheral blood lymphocyte mRNA to isolate cDNA clones that include the carboxyl-terminal coding region of human CD6. The nucleotide sequence of the longest human cDNA clone, CD6-PB1, predicts a protein of 668 amino acids with a 244-amino acid cytoplasmic domain similar in size to and possessing 71.5% amino acid sequence identity with the cytoplasmic domain of mouse CD6. This previously unrecognized 244-amino acid cytoplasmic domain does not have significant homology to any other known protein (except mouse CD6), but does possess two proline-rich motifs containing the SH3 domain-binding consensus sequence, a serine-threonine-rich motif repeated three times, three protein kinase C phosphorylation-site motifs, and 10 casein kinase-2 phosphorylation-site motifs. These sequences are likely to play a role in the ability of CD6-specific monoclonal antibodies to stimulate T cell proliferation. Full-length CD6 cDNA containing this cytoplasmic domain sequence encodes a monomeric 105/130-kDa protein that can be immunoprecipitated from the surface of transfected cells and comigrates upon SDS-PAGE with wild-type CD6 immunoprecipitated from PBL. We also isolated two alternatively spliced forms of human CD6 cDNA lacking sequences encoding membrane-proximal regions of the cytoplasmic domain which maintain the same reading frame as CD6-PB1. The short cytoplasmic domain of the previously reported human CD6-15 cDNA clone results from a deletion of a 20-bp segment through use of an alternative 3' splice site, resulting in a frame shift and premature termination of translation relative to the clones we have isolated. These data demonstrate that human CD6 possesses a large cytoplasmic domain containing sequence motifs that are likely to be involved in signal transduction upon stimulation of T cells through CD6 ligation.

  View details for Web of Science ID A1995TB37100007

  View details for PubMedID 7589069

• STRUCTURE OF THE MOUSE CD72 (LYB-2) GENE AND ITS ALTERNATIVELY SPLICED TRANSCRIPTS JOURNAL OF IMMUNOLOGY Ying, H., Nakayama, E., Robinson, W. H., Parnes, J. R. 1995; 154 (6): 2743-2752

  Abstract

  The complete sequence of the CD72 gene from the C57L mouse, including the 5' and 3' flanking sequences, is reported. The gene spans 6830 base pairs and includes nine exons surrounding eight introns. It does not have an obvious TATAA box, so it belongs to a group of genes with TATA-less promoters that are regulated during mammalian immunodifferentiation. cDNA sequence comparisons among CD72a, CD72b, and CD72c alleles have demonstrated two distinct seven amino acid insertion/deletions among these allelic variants. Based on our genomic sequence studies as well as PCR analyses, we found that different strains of mice can alternatively or exclusively use either of two AG sites surrounding the 21-bp insertion/deletions as 3' splice sites in an allele-specific manner. Other alternative splicing events, such as exon skipping, also contribute to CD72 polymorphism. In mouse splenic B cells there are allele-specific distributions of CD72 mRNAs that contain sequences from both exon 3 and exon 4, from either exon 3 or exon 4, or from neither exon 3 nor 4. It is unclear what the in vivo function might be of the proteins encoded by the mRNA forms lacking these exon sequences.

  View details for Web of Science ID A1995QL08400021

  View details for PubMedID 7876545

• BIOCHEMICAL IDENTITY OF THE MOUSE LY-19.2 AND LY-32.2 ALLOANTIGENS WITH THE B-CELL DIFFERENTIATION ANTIGEN LYB-2/CD72 JOURNAL OF IMMUNOLOGY Robinson, W. H., LANDOLFI, M. M., Schafer, H., Parnes, J. R. 1993; 151 (9): 4764-4772

  Abstract

  Lyb-2/CD72 is a 45-kDa mouse B cell surface protein that binds CD5 (Ly-1) and has been shown to induce B cell proliferation upon mAb binding. The serologically defined Ly-19.2 and Ly-32.2 lymphocyte alloantigens have mouse strain distribution patterns similar to that of the Lyb-2/CD72 alleles and map to the same region on chromosome 4 as Lyb-2/CD72. Our recent isolation of the Lyb-2a, -2b, and -2c cDNA has enabled us in this report to examine the relationship between Ly-19, Ly-32, and Lyb-2/CD72. A rat T cell line transfected with a mouse Lyb-2a cDNA is recognized by Ly-19.2-specific mAb, whereas transfectants expressing the Lyb-2b cDNA are recognized by both Ly-19.2 and Ly-32.2-specific mAb. Cell surface iodination immunoprecipitation analysis from Lyb-2a cDNA transfectants using Lyb-2a- and Ly-19.2-specific mAb as well as from Lyb-2b cDNA transfectants using Lyb-2b-, Ly-19.2-, and Ly-32.2-specific Ab, produced immunoprecipitates containing comigrating 45-kDa polypeptides. Preclearing studies with these transfectants indicate that the immunoprecipitated proteins represent the same polypeptide chain. These results demonstrate that the mouse Ly-19.2 and Ly-32.2 alloantigens are in fact the B cell differentiation Ag Lyb-2/CD72.

  View details for Web of Science ID A1993MD34900035

  View details for PubMedID 8409435

• EXTENSIVE POLYMORPHISM IN THE EXTRACELLULAR DOMAIN OF THE MOUSE B-CELL DIFFERENTIATION ANTIGEN LYB-2/CD72 JOURNAL OF IMMUNOLOGY Robinson, W. H., Ying, H., Miceli, M. C., Parnes, J. R. 1992; 149 (3): 880-886

  Abstract

  Lyb-2/CD72 is a 45-kDa mouse B cell surface protein that binds CD5 and has been shown to play a role in B cell proliferation and differentiation. Using the polymerase chain reaction we have isolated and sequenced cDNA clones encoding the serologically defined mouse Lyb-2a, Lyb-2b, and Lyb-2c alleles. We confirmed that our full length cDNA clones encode the Lyb-2a, -2b, and -2c alleles, respectively, by transfecting the isolated Lyb-2/CD72 cDNA clones into L cells and demonstrating that the transfectants bind only the appropriate allele specific anti-Lyb-2/CD72 antibodies. Sequence comparisons demonstrate that the Lyb-2/CD72 allels are highly conserved in their cytoplasmic and transmembrane domains but exhibit a high degree of polymorphism in their extracellular domains. This polymorphism in the extracellular region involves amino acid substitutions at a minimum of 20 residues and is concentrated primarily in the membrane distal region. cDNA sequence comparisons also demonstrate two distinct seven amino acid insertion/deletions among these allelic variants. A form of Lyb-2b cDNA lacking the sequence encoding the transmembrane region was isolated from a C57B1/6 mouse and a CH12.LX subline. The Lyb-2/CD72 PCR products from mRNA of mice expressing Lyb-2a and Lyb-2c contain a DNA fragment that corresponds in size to the transmembraneless form, suggesting that these mouse strains also express this mRNA.

  View details for Web of Science ID A1992JF47400019

  View details for PubMedID 1634777

• SYNTHESIS OF VIRAL-SPECIFIC RIBONUCLEIC ACID IN RUBELLA VIRUS-INFECTED CELLS JOURNAL OF VIROLOGY Wong, K. T., Robinson, W. S., Merigan, T. C. 1969; 4 (6): 901-?

  Abstract

  Monolayers of BHK-21/W1-2 cells were pulsed with (3)H-uridine at different times after infection with rubella virus, and viral-specific cytoplasmic ribonucleic acid species were demonstrated.

  View details for Web of Science ID A1969E851000014

  View details for PubMedID 16789124

  View details for PubMedCentralID PMC375955