{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T12:51:03Z","timestamp":1788612663321,"version":"build-2803163510"},"reference-count":35,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T00:00:00Z","timestamp":1740528000000},"content-version":"vor","delay-in-days":2370,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Rheumatoid arthritis (RA) is characterized by formation of synovial ectopic lymphoid structures (ELS) supporting B cell autoreactivity toward locally generated citrullinated (cit) antigens, including those contained in neutrophil extracellular traps (NETs). However, only a minority of RA-rmAbs from B cells isolated from ELS+ RA tissues react against NETs. Thus, alternative cellular sources of other potential autoantigens targeted by locally differentiated B cells remain undefined. RA fibroblast\u2013like synoviocytes (FLS) have been implicated in the release of RA-associated autoantigens. In this study, we aimed to define stromal-derived autoantigens from RA-FLS targeted by RA-rmAbs. Seventy-one RA-rmAbs were screened toward RA-FLS by living-cell immunofluorescence (IF). Western blotting was used to identify potential autoantigens from RA-FLS protein extracts. Putative candidates were validated using colocalization immunofluorescence confocal microscopy, ELISA, immunoprecipitation assay, and surface plasmon resonance on unmodified\/cit proteins. Serum immunoreactivity was tested in anti-citrullinated peptide\/protein Abs (ACPA)+ versus ACPA\u2212 RA patients. Ten out of 71 RA-rmAbs showed clear reactivity toward RA-FLS in immunofluorescence with no binding to NETs. One stromal-reactive RA-rmAb (RA057\/11.89.1) decorated a \u223c58-kDa band that mass spectrometry and Western blotting with a commercial Ab identified as calreticulin (CRT). Confocal microscopy demonstrated significant cellular colocalization between anti-CRT RA057\/11.89.1 in RA-FLS. RA057\/11.89.1 was able to immunoprecipitate rCRT. Deimination of CRT to cit-CRT moderately increased RA057\/11.89.1 immunoreactivity. cit-CRT displayed increased blocking capacity compared with unmodified CRT in competitive binding assays. Finally, anti\u2013cit-CRT Abs were preferentially detected in ACPA+ versus ACPA\u2212 RA sera. We identified a synovial B cell\u2013derived RA-rmAb locally differentiated within the ELS+ RA synovium reacting toward CRT, a putative novel autoantigen recently described in RA patients, suggesting that FLS-derived CRT may contribute to fuel the local autoimmune response.<\/jats:p>","DOI":"10.4049\/jimmunol.1800346","type":"journal-article","created":{"date-parts":[[2018,7,25]],"date-time":"2018-07-25T14:25:19Z","timestamp":1532528719000},"page":"1373-1381","source":"Crossref","is-referenced-by-count":13,"title":["Characterization of a Synovial B Cell\u2013Derived Recombinant Monoclonal Antibody Targeting Stromal Calreticulin in the Rheumatoid Joints"],"prefix":"10.1093","volume":"201","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2092-6778","authenticated-orcid":false,"given":"Elisa","family":"Corsiero","sequence":"first","affiliation":[{"name":"Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , London EC1M 6BQ ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6412-2619","authenticated-orcid":false,"given":"Lucas","family":"Jagemann","sequence":"additional","affiliation":[{"name":"Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , London EC1M 6BQ ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2068-3331","authenticated-orcid":false,"given":"Mauro","family":"Perretti","sequence":"additional","affiliation":[{"name":"Centre for Biochemical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , London EC1M 6BQ ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1326-5051","authenticated-orcid":false,"given":"Costantino","family":"Pitzalis","sequence":"additional","affiliation":[{"name":"Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , London EC1M 6BQ ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele","family":"Bombardieri","sequence":"additional","affiliation":[{"name":"Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , London EC1M 6BQ ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2018,9,1]]},"reference":[{"key":"2025030417572265900_r1","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1371\/journal.pmed.0060001","article-title":"Ectopic lymphoid structures support ongoing production of class-switched autoantibodies in rheumatoid synovium","volume":"6","author":"Humby","year":"2009","journal-title":"PLoS Med."},{"key":"2025030417572265900_r2","doi-asserted-by":"crossref","first-page":"1866","DOI":"10.1136\/annrheumdis-2015-208356","article-title":"Single cell cloning and recombinant monoclonal antibodies generation from RA synovial B cells reveal frequent targeting of citrullinated histones of NETs","volume":"75","author":"Corsiero","year":"2016","journal-title":"Ann. Rheum. Dis."},{"key":"2025030417572265900_r3","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1038\/nrrheum.2016.217","article-title":"Ectopic lymphoid neogenesis in rheumatic autoimmune diseases","volume":"13","author":"Bombardieri","year":"2017","journal-title":"Nat. Rev. Rheumatol."},{"key":"2025030417572265900_r4","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1002\/art.27767","article-title":"V-region gene analysis of locally defined synovial B and plasma cells reveals selected B cell expansion and accumulation of plasma cell clones in rheumatoid arthritis","volume":"63","author":"Scheel","year":"2011","journal-title":"Arthritis Rheum."},{"key":"2025030417572265900_r5","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/S0002-9440(10)61762-2","article-title":"Thyroid autoimmune disease: demonstration of thyroid antigen-specific B cells and recombination-activating gene expression in chemokine-containing active intrathyroidal germinal centers","volume":"159","author":"Armengol","year":"2001","journal-title":"Am. J. Pathol."},{"key":"2025030417572265900_r6","doi-asserted-by":"crossref","first-page":"3187","DOI":"10.1002\/art.11311","article-title":"Cellular basis of ectopic germinal center formation and autoantibody production in the target organ of patients with Sj\u00f6gren\u2019s syndrome","volume":"48","author":"Salomonsson","year":"2003","journal-title":"Arthritis Rheum."},{"key":"2025030417572265900_r7","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.jaut.2010.07.004","article-title":"Somatic hypermutation and antigen-driven selection of B cells are altered in autoimmune diseases","volume":"35","author":"Zuckerman","year":"2010","journal-title":"J. Autoimmun."},{"key":"2025030417572265900_r8","doi-asserted-by":"crossref","first-page":"430","DOI":"10.3389\/fimmu.2016.00430","article-title":"Ectopic lymphoid structures: powerhouse of autoimmunity","volume":"7","author":"Corsiero","year":"2016","journal-title":"Front. Immunol."},{"key":"2025030417572265900_r9","doi-asserted-by":"crossref","first-page":"2205","DOI":"10.1056\/NEJMra1004965","article-title":"The pathogenesis of rheumatoid arthritis","volume":"365","author":"McInnes","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"2025030417572265900_r10","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/rheumatology\/kel065","article-title":"Synovial fibroblasts: key players in rheumatoid arthritis","volume":"45","author":"Huber","year":"2006","journal-title":"Rheumatology (Oxford)"},{"key":"2025030417572265900_r11","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1136\/ard.2011.150219","article-title":"A BAFF\/APRIL-dependent TLR3-stimulated pathway enhances the capacity of rheumatoid synovial fibroblasts to induce AID expression and Ig class-switching in B cells","volume":"70","author":"Bombardieri","year":"2011","journal-title":"Ann. Rheum. 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Dis."},{"key":"2025030417572265900_r15","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/s00011-012-0572-1","article-title":"Reduced apoptosis correlates with enhanced autophagy in synovial tissues of rheumatoid arthritis","volume":"62","author":"Xu","year":"2013","journal-title":"Inflamm. Res."},{"key":"2025030417572265900_r16","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1186\/ar4074","article-title":"Epigenetic contributions in the development of rheumatoid arthritis","volume":"14","author":"Klein","year":"2012","journal-title":"Arthritis Res. 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Rheumatol."},{"key":"2025030417572265900_r21","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1002\/cyto.a.20896","article-title":"Quantifying colocalization by correlation: the Pearson correlation coefficient is superior to the Mander\u2019s overlap coefficient","volume":"77","author":"Adler","year":"2010","journal-title":"Cytometry A"},{"key":"2025030417572265900_r22","doi-asserted-by":"crossref","first-page":"4561","DOI":"10.4049\/jimmunol.1000536","article-title":"Functional analysis of recombinant calreticulin fragment 39-272: implications for immunobiological activities of calreticulin in health and disease","volume":"185","author":"Hong","year":"2010","journal-title":"J. Immunol."},{"key":"2025030417572265900_r23","doi-asserted-by":"crossref","first-page":"R161","DOI":"10.1186\/ar1153","article-title":"The synovial proteome: analysis of fibroblast-like synoviocytes","volume":"6","author":"Dasuri","year":"2004","journal-title":"Arthritis Res. 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Sci."},{"key":"2025030417572265900_r26","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.it.2012.08.002","article-title":"Calreticulin in the immune system: ins and outs","volume":"34","author":"Raghavan","year":"2013","journal-title":"Trends Immunol."},{"key":"2025030417572265900_r27","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1111\/cei.12411","article-title":"Calreticulin promotes angiogenesis via activating nitric oxide signalling pathway in rheumatoid arthritis","volume":"178","author":"Ding","year":"2014","journal-title":"Clin. Exp. Immunol."},{"key":"2025030417572265900_r28","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1007\/s10875-013-9885-2","article-title":"Serum levels of calreticulin in correlation with disease activity in patients with rheumatoid arthritis","volume":"33","author":"Ni","year":"2013","journal-title":"J. Clin. 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An approach to understanding the molecular genetics of susceptibility to rheumatoid arthritis","volume":"30","author":"Gregersen","year":"1987","journal-title":"Arthritis Rheum."},{"key":"2025030417572265900_r34","doi-asserted-by":"crossref","first-page":"R1386","DOI":"10.1186\/ar1837","article-title":"The impact of HLA-DRB1 genes on extra-articular disease manifestations in rheumatoid arthritis","volume":"7","author":"Turesson","year":"2005","journal-title":"Arthritis Res. Ther."},{"key":"2025030417572265900_r35","doi-asserted-by":"crossref","first-page":"6359","DOI":"10.4049\/jimmunol.179.9.6359","article-title":"The rheumatoid arthritis shared epitope triggers innate immune signaling via cell surface calreticulin","volume":"179","author":"Ling","year":"2007","journal-title":"J. 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