{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T09:14:19Z","timestamp":1773306859878,"version":"3.50.1"},"reference-count":27,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2017,4,26]],"date-time":"2017-04-26T00:00:00Z","timestamp":1493164800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Arthritis &amp; Rheumatology"],"published-print":{"date-parts":[[2017,5]]},"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>Most patients with rheumatoid arthritis (RA) harbor antibodies to citrullinated autoantigens such as citrullinated fibrinogen. Two isoforms of peptidylarginine deiminase (PAD), PAD type 2 (PAD2) and PAD4, which catalyze citrullination with different substrate specificities, can be detected in the synovium of RA patients. This study was undertaken to determine whether RA antibodies preferentially bind PAD2\u2010 or PAD4\u2010citrullinated fibrinogen.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>RA patient and normal donor plasma specimens were tested for binding to PAD2\u2010 or PAD4\u2010citrullinated fibrinogen, native fibrinogen, or citrullinated fibrinogen peptides in various dilutions by enzyme\u2010linked immunosorbent assay (ELISA) and Western blotting. Bands corresponding to masses demonstrating RA antibody reactivity by Western blotting were excised and analyzed by mass spectrometry.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>At low antibody titers (1:40 and 1:100), there was no significant difference between RA antibody reactivity to PAD2\u2010 and PAD4\u2010citrullinated fibrinogen. When plasma was further diluted to 1:250 and 1:1,000, RA patient plasma bound PAD4\u2010citrullinated fibrinogen significantly more than PAD2\u2010citrullinated fibrinogen, as measured by ELISA and Western blotting. An increased antibody titer was associated with increased avidity for both PAD2\u2010 and PAD4\u2010citrullinated fibrinogen. Both enzymes hypercitrullinated fibrinogen, but PAD4 citrullinated arginines more intermittently, generating a mix of citrullinated and noncitrullinated arginines. Peptide ELISA and preadsorption assays confirmed that the region of intermittent citrullination accounts for the majority of RA antibody binding to the \u03b2\u2010chain of citrullinated fibrinogen.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>At high titers, RA antibodies preferentially bind fibrinogen modified by PAD4, because intermittent citrullination offers a more diverse assortment of citrullinated epitopes.<\/jats:p><\/jats:sec>","DOI":"10.1002\/art.40035","type":"journal-article","created":{"date-parts":[[2016,12,28]],"date-time":"2016-12-28T11:52:59Z","timestamp":1482925979000},"page":"986-995","source":"Crossref","is-referenced-by-count":33,"title":["High\u2010Titer Rheumatoid Arthritis Antibodies Preferentially Bind Fibrinogen Citrullinated by Peptidylarginine Deiminase 4"],"prefix":"10.1002","volume":"69","author":[{"given":"Nathalie E.","family":"Blach\u00e8re","sequence":"first","affiliation":[{"name":"Rockefeller University and Howard Hughes Medical Institute New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salina","family":"Parveen","sequence":"additional","affiliation":[{"name":"Rockefeller University New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mayu O.","family":"Frank","sequence":"additional","affiliation":[{"name":"Rockefeller University and New York Genome Center New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian D.","family":"Dill","sequence":"additional","affiliation":[{"name":"Rockefeller University New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henrik","family":"Molina","sequence":"additional","affiliation":[{"name":"Rockefeller University New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dana E.","family":"Orange","sequence":"additional","affiliation":[{"name":"Rockefeller University, Hospital for Special Surgery, and New York Genome Center New York New York"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,4,26]]},"reference":[{"issue":"1","key":"e_1_2_7_2_1","first-page":"S17","article-title":"Anti\u2010CCP antibodies: the new rheumatoid factor in the serology of rheumatoid arthritis","volume":"3","author":"Venrooij WJ","year":"2004","journal-title":"Autoimmun Rev"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1136\/ard.2008.091355"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/art.23763"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/art.25036"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/ar2011"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1001620"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/art.22983"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3005580"},{"key":"e_1_2_7_10_1","first-page":"3135","article-title":"Release of active peptidyl arginine deiminases by neutrophils can explain production of extracellular citrullinated autoantigens in rheumatoid arthritis synovial fluid. 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