{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T05:20:32Z","timestamp":1770960032453,"version":"3.50.1"},"reference-count":19,"publisher":"Oxford University Press (OUP)","issue":"9","license":[{"start":{"date-parts":[[2020,5,1]],"date-time":"2020-05-01T00:00:00Z","timestamp":1588291200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100012041","name":"Versus Arthritis","doi-asserted-by":"publisher","award":["20858"],"award-info":[{"award-number":["20858"]}],"id":[{"id":"10.13039\/501100012041","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,5,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>We previously showed that anti\u2013neutrophil extracellular trap (NET) rheumatoid arthritis (RA)-rmAbs derived from CD19+ B cells within RA human synovial tissues frequently react against NETs. In this study, we aimed to characterize the importance of affinity maturation via somatic hypermutation (SHM) within the Ig variable H (VH) and variable L (VL) chains and Fab\u2013N-linked glycosylation in RA synovial B cell clones reactive to NETs and NET-derived Ags such as citrullinated histones. Selected anti-NET RA-rmAbs derived from synovial RA CD19+ B cells were subjected to overlap-PCR to generate germline (GL; VH and VL reverted into GL), hybrid clones (VH\/VL region reverted into GL), and N-glycosylation mutants (N\u2192Q) and analyzed for anti-NETs and citrullinated histones (cit-H2B) immunoreactivity. Anti-NET\/cit-H2B immunoreactivity of selected RA-rmAbs was abrogated in the VH and VL GL counterpart. In RA B cell hybrid clone RA015\/11.88 and RA056\/11.23.2, NET and\/or cit-H2B immunoreactivity was solely dependent on SHM in the IgVH region whereas RA B cell hybrid clone RA015\/11.91 required affinity maturation of both VH and VL for efficient binding to cit-H2B. In 7\/80 RA-rmAb, SHM resulted in ex novo N-glycosylation sites in VH and\/or VL regions. Removal of Fab-linked glycans in RA056\/11.23.2 in the N-mutant counterpart resulted in 90% reduction in immunoreactivity to cit-H2B. Thus, SHM in the IgVH and\/or VL regions of RA synovial B cells is necessary for the immunoreactivity to NET-Ags. Fab\u2013N-linked\u2013glycosylation introduction sites are observed in a minority of anti-NET B cell clones but can strongly influence NET-Ag binding.<\/jats:p>","DOI":"10.4049\/jimmunol.1901457","type":"journal-article","created":{"date-parts":[[2020,3,27]],"date-time":"2020-03-27T22:30:47Z","timestamp":1585348247000},"page":"2374-2379","source":"Crossref","is-referenced-by-count":8,"title":["H and L Chain Affinity Maturation and\/or Fab <i>N<\/i>-Glycosylation Influence Immunoreactivity toward Neutrophil Extracellular Trap Antigens in Rheumatoid Arthritis Synovial B Cell Clones"],"prefix":"10.1093","volume":"204","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":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2794-9131","authenticated-orcid":false,"given":"Emanuela","family":"Carlotti","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":[{"role":"author","vocabulary":"crossref"}]},{"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":[{"role":"author","vocabulary":"crossref"}]},{"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":[{"role":"author","vocabulary":"crossref"}]},{"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":[{"role":"author","vocabulary":"crossref"}]},{"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":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2020,5,1]]},"reference":[{"key":"2025030417511635100_r1","doi-asserted-by":"crossref","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":"2025030417511635100_r2","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":"2025030417511635100_r3","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. 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