{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T02:44:42Z","timestamp":1792032282521,"version":"4.4.0"},"reference-count":24,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,6,6]],"date-time":"2021-06-06T00:00:00Z","timestamp":1622937600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,6,6]],"date-time":"2021-06-06T00:00:00Z","timestamp":1622937600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR066712"],"award-info":[{"award-number":["AR066712"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006108","name":"National Center for Advancing Translational Sciences","doi-asserted-by":"publisher","award":["UL1 TR001082-04"],"award-info":[{"award-number":["UL1 TR001082-04"]}],"id":[{"id":"10.13039\/100006108","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["U01 AI101981"],"award-info":[{"award-number":["U01 AI101981"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007155","name":"Walter S. and Lucienne Driskill Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007155","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Arthritis Res Ther"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p>Mucosal sites are hypothesized to play a role in the development of rheumatoid arthritis (RA). Since serum anti-peptidylarginine deiminase (PAD)4 antibodies, including a subset that cross-react with PAD3 (PAD3\/4), are specific for RA and associate with severe disease, we sought to examine whether anti-PAD4 and anti-PAD3\/4 antibodies were present in the lung and oral mucosa of subjects with RA and \u201cat-risk\u201d for RA.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>We included 37 RA, 25 healthy control, and 46 subjects \u201cat-risk\u201d for RA based on familial RA and\/or serum anti-citrullinated protein antibody (ACPA) positivity. Paired serum, sputum, and saliva were evaluated for anti-PAD4 and anti-PAD3\/4 using immunoprecipitation and ACPA using ELISA. Immunoglobulins (Ig) were purified from representative samples, and their effect on citrullination of histone H3 by recombinant human PAD4 was measured by anti-citH3 immunoblot.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Anti-PAD4 antibodies were detected in the serum of 6\/37 (16.2%), sputum of 3\/37 (8.1%), and saliva of 3\/33 (9.1%) RA subjects and in the serum and sputum of 1\/46 (2.2%) at-risk subjects. None of the healthy controls had anti-PAD4 antibodies at any site. Serum, sputum, and salivary anti-PAD4 antibodies were more prevalent in RA subjects with RA duration &gt;2 years. Purified antibodies from representative anti-PAD4-positive and anti-PAD3\/4-positive sputum were primarily of the IgA isotype and able to increase PAD4 enzymatic activity.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Anti-PAD4 antibodies are present in the sputum and saliva of a portion of RA patients and are infrequent in at-risk subjects. Importantly, the ability of anti-PAD4, and particularly anti-PAD3\/4, antibodies in the sputum to enhance PAD4 enzymatic activity suggests that anti-PAD4 may play an active role in the RA lung.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s13075-021-02528-5","type":"journal-article","created":{"date-parts":[[2021,6,6]],"date-time":"2021-06-06T22:04:47Z","timestamp":1623017087000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Anti-peptidylarginine deiminase-4 antibodies at mucosal sites can activate peptidylarginine deiminase-4 enzyme activity in rheumatoid arthritis"],"prefix":"10.1186","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2563-5155","authenticated-orcid":false,"given":"M. Kristen","family":"Demoruelle","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan L.","family":"Davis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley","family":"Visser","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johnny","family":"Hoang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jill M.","family":"Norris","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V. Michael","family":"Holers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin D.","family":"Deane","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erika","family":"Darrah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,6,6]]},"reference":[{"issue":"7","key":"2528_CR1","doi-asserted-by":"publisher","first-page":"700","DOI":"10.2174\/1389450116666150202160954","volume":"16","author":"EE Witalison","year":"2015","unstructured":"Witalison EE, Thompson PR, Hofseth LJ. Protein arginine deiminases and associated citrullination: physiological functions and diseases associated with dysregulation. Curr Drug Targets. 2015;16(7):700\u201310. https:\/\/doi.org\/10.2174\/1389450116666150202160954.","journal-title":"Curr Drug Targets."},{"issue":"186","key":"2528_CR2","doi-asserted-by":"publisher","first-page":"186ra65","DOI":"10.1126\/scitranslmed.3005370","volume":"5","author":"E Darrah","year":"2013","unstructured":"Darrah E, Giles JT, Ols ML, Bull HG, Andrade F, Rosen A. Erosive rheumatoid arthritis is associated with antibodies that activate PAD4 by increasing calcium sensitivity. Sci Transl Med. 2013;5(186):186ra65.","journal-title":"Sci Transl Med"},{"issue":"6","key":"2528_CR3","doi-asserted-by":"publisher","first-page":"e98794","DOI":"10.1371\/journal.pone.0098794","volume":"9","author":"JT Giles","year":"2014","unstructured":"Giles JT, Darrah E, Danoff S, Johnson C, Andrade F, Rosen A, et al. Association of cross-reactive antibodies targeting peptidyl-arginine deiminase 3 and 4 with rheumatoid arthritis-associated interstitial lung disease. PLoS One. 2014;9(6):e98794. https:\/\/doi.org\/10.1371\/journal.pone.0098794.","journal-title":"PLoS One."},{"issue":"7","key":"2528_CR4","doi-asserted-by":"publisher","first-page":"1958","DOI":"10.1002\/art.23596","volume":"58","author":"ML Harris","year":"2008","unstructured":"Harris ML, Darrah E, Lam GK, Bartlett SJ, Giles JT, Grant AV, et al. Association of autoimmunity to peptidyl arginine deiminase type 4 with genotype and disease severity in rheumatoid arthritis. Arthritis Rheum. 2008;58(7):1958\u201367. https:\/\/doi.org\/10.1002\/art.23596.","journal-title":"Arthritis Rheum."},{"issue":"3","key":"2528_CR5","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1136\/ard.2007.080267","volume":"67","author":"EH Halvorsen","year":"2008","unstructured":"Halvorsen EH, Pollmann S, Gilboe IM, van der Heijde D, Landewe R, Odegard S, et al. Serum IgG antibodies to peptidylarginine deiminase 4 in rheumatoid arthritis and associations with disease severity. Ann Rheum Dis. 2008;67(3):414\u20137. https:\/\/doi.org\/10.1136\/ard.2007.080267.","journal-title":"Ann Rheum Dis."},{"issue":"6","key":"2528_CR6","doi-asserted-by":"publisher","first-page":"903","DOI":"10.1002\/art.41189","volume":"72","author":"I Auger","year":"2020","unstructured":"Auger I, Balandraud N, Massy E, Hemon MF, Peen E, Arnoux F, et al. Peptidylarginine deiminase autoimmunity and the development of anti-citrullinated protein antibody in rheumatoid arthritis: the hapten-carrier model. Arthritis Rheumatol. 2020;72(6):903\u201311. https:\/\/doi.org\/10.1002\/art.41189.","journal-title":"Arthritis Rheumatol."},{"issue":"6","key":"2528_CR7","first-page":"969","volume":"35","author":"J Zhao","year":"2008","unstructured":"Zhao J, Zhao Y, He J, Jia R, Li Z. Prevalence and significance of anti-peptidylarginine deiminase 4 antibodies in rheumatoid arthritis. J Rheumatol. 2008;35(6):969\u201374.","journal-title":"J Rheumatol."},{"issue":"3","key":"2528_CR8","doi-asserted-by":"publisher","first-page":"434","DOI":"10.1136\/annrheumdis-2018-213818","volume":"78","author":"L Martinez-Prat","year":"2019","unstructured":"Martinez-Prat L, Lucia D, Ibarra C, Mahler M, Dervieux T. Antibodies targeting protein-arginine deiminase 4 (PAD4) demonstrate diagnostic value in rheumatoid arthritis. Ann Rheum Dis. 2019;78(3):434\u20136. https:\/\/doi.org\/10.1136\/annrheumdis-2018-213818.","journal-title":"Ann Rheum Dis."},{"issue":"9","key":"2528_CR9","doi-asserted-by":"publisher","first-page":"2633","DOI":"10.1002\/art.27570","volume":"62","author":"JR Kolfenbach","year":"2010","unstructured":"Kolfenbach JR, Deane KD, Derber LA, O\u2019Donnell CI, Gilliland WR, Edison JD, et al. Autoimmunity to peptidyl arginine deiminase type 4 precedes clinical onset of rheumatoid arthritis. Arthritis Rheum. 2010;62(9):2633\u20139. https:\/\/doi.org\/10.1002\/art.27570.","journal-title":"Arthritis Rheum."},{"issue":"9","key":"2528_CR10","doi-asserted-by":"publisher","first-page":"542","DOI":"10.1038\/s41584-018-0070-0","volume":"14","author":"VM Holers","year":"2018","unstructured":"Holers VM, Demoruelle MK, Kuhn KA, Buckner JH, Robinson WH, Okamoto Y, et al. Rheumatoid arthritis and the mucosal origins hypothesis: protection turns to destruction. Nat Rev Rheumatol. 2018;14(9):542\u201357. https:\/\/doi.org\/10.1038\/s41584-018-0070-0.","journal-title":"Nat Rev Rheumatol."},{"key":"2528_CR11","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1136\/annrheumdis-2020-216972","volume":"80","author":"Z Cheng","year":"2020","unstructured":"Cheng Z, Do T, Mankia K, Meade J, Hunt L, Clerehugh V, et al. Dysbiosis in the oral microbiomes of anti-CCP positive individuals at risk of developing rheumatoid arthritis. Ann Rheum Dis. 2020;80:162\u20138.","journal-title":"Ann Rheum Dis"},{"issue":"1","key":"2528_CR12","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.berh.2017.08.002","volume":"31","author":"V Joshua","year":"2017","unstructured":"Joshua V, Chatzidionisyou K, Catrina AI. Role of the lung in individuals at risk of rheumatoid arthritis. Best Pract Res Clin Rheumatol. 2017;31(1):31\u201341. https:\/\/doi.org\/10.1016\/j.berh.2017.08.002.","journal-title":"Best Pract Res Clin Rheumatol."},{"issue":"6","key":"2528_CR13","doi-asserted-by":"publisher","first-page":"1165","DOI":"10.1002\/art.40066","volume":"69","author":"MK Demoruelle","year":"2017","unstructured":"Demoruelle MK, Harrall KK, Ho L, Purmalek MM, Seto NL, Rothfuss HM, et al. Anti-citrullinated protein antibodies are associated with neutrophil extracellular traps in the sputum in relatives of rheumatoid arthritis patients. Arthritis Rheumatol. 2017;69(6):1165\u201375. https:\/\/doi.org\/10.1002\/art.40066.","journal-title":"Arthritis Rheumatol."},{"issue":"10","key":"2528_CR14","doi-asserted-by":"publisher","first-page":"2545","DOI":"10.1002\/art.38066","volume":"65","author":"VC Willis","year":"2013","unstructured":"Willis VC, Demoruelle MK, Derber LA, Chartier-Logan CJ, Parish MC, Pedraza IF, et al. Sputum autoantibodies in patients with established rheumatoid arthritis and subjects at risk of future clinically apparent disease. Arthritis Rheum. 2013;65(10):2545\u201354. https:\/\/doi.org\/10.1002\/art.38066.","journal-title":"Arthritis Rheum."},{"issue":"1","key":"2528_CR15","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1186\/s13075-019-1808-z","volume":"21","author":"B Rodriguez-Lozano","year":"2019","unstructured":"Rodriguez-Lozano B, Gonzalez-Febles J, Garnier-Rodriguez JL, Dadlani S, Bustabad-Reyes S, Sanz M, et al. Association between severity of periodontitis and clinical activity in rheumatoid arthritis patients: a case-control study. Arthritis Res Ther. 2019;21(1):27. https:\/\/doi.org\/10.1186\/s13075-019-1808-z.","journal-title":"Arthritis Res Ther."},{"issue":"1","key":"2528_CR16","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1186\/s40168-016-0206-x","volume":"4","author":"JU Scher","year":"2016","unstructured":"Scher JU, Joshua V, Artacho A, Abdollahi-Roodsaz S, Ockinger J, Kullberg S, et al. The lung microbiota in early rheumatoid arthritis and autoimmunity. Microbiome. 2016;4(1):60. https:\/\/doi.org\/10.1186\/s40168-016-0206-x.","journal-title":"Microbiome."},{"issue":"12","key":"2528_CR17","doi-asserted-by":"publisher","first-page":"1735","DOI":"10.1002\/art.24833","volume":"61","author":"JR Kolfenbach","year":"2009","unstructured":"Kolfenbach JR, Deane KD, Derber LA, O\u2019Donnell C, Weisman MH, Buckner JH, et al. A prospective approach to investigating the natural history of preclinical rheumatoid arthritis (RA) using first-degree relatives of probands with RA. Arthritis Rheum. 2009;61(12):1735\u201342. https:\/\/doi.org\/10.1002\/art.24833.","journal-title":"Arthritis Rheum."},{"issue":"6","key":"2528_CR18","doi-asserted-by":"publisher","first-page":"1630","DOI":"10.1002\/art.27439","volume":"62","author":"F Andrade","year":"2010","unstructured":"Andrade F, Darrah E, Gucek M, Cole RN, Rosen A, Zhu X. Autocitrullination of human peptidyl arginine deiminase type 4 regulates protein citrullination during cell activation. Arthritis Rheum. 2010;62(6):1630\u201340. https:\/\/doi.org\/10.1002\/art.27439.","journal-title":"Arthritis Rheum."},{"issue":"1","key":"2528_CR19","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1002\/art.27230","volume":"62","author":"I Auger","year":"2010","unstructured":"Auger I, Martin M, Balandraud N, Roudier J. Rheumatoid arthritis-specific autoantibodies to peptidyl arginine deiminase type 4 inhibit citrullination of fibrinogen. Arthritis Rheum. 2010;62(1):126\u201331. https:\/\/doi.org\/10.1002\/art.27230.","journal-title":"Arthritis Rheum."},{"issue":"9","key":"2528_CR20","doi-asserted-by":"publisher","first-page":"1722","DOI":"10.1136\/annrheumdis-2015-208216","volume":"75","author":"G Reynisdottir","year":"2016","unstructured":"Reynisdottir G, Olsen H, Joshua V, Engstrom M, Forsslund H, Karimi R, et al. Signs of immune activation and local inflammation are present in the bronchial tissue of patients with untreated early rheumatoid arthritis. Ann Rheum Dis. 2016;75(9):1722\u20137. https:\/\/doi.org\/10.1136\/annrheumdis-2015-208216.","journal-title":"Ann Rheum Dis."},{"issue":"2","key":"2528_CR21","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1111\/cei.12677","volume":"182","author":"Z Reyes-Castillo","year":"2015","unstructured":"Reyes-Castillo Z, Palafox-Sanchez CA, Parra-Rojas I, Martinez-Bonilla GE, del Toro-Arreola S, Ramirez-Duenas MG, et al. Comparative analysis of autoantibodies targeting peptidylarginine deiminase type 4, mutated citrullinated vimentin and cyclic citrullinated peptides in rheumatoid arthritis: associations with cytokine profiles, clinical and genetic features. Clin Exp Immunol. 2015;182(2):119\u201331. https:\/\/doi.org\/10.1111\/cei.12677.","journal-title":"Clin Exp Immunol."},{"key":"2528_CR22","doi-asserted-by":"publisher","first-page":"2696","DOI":"10.3389\/fimmu.2018.02696","volume":"9","author":"E Darrah","year":"2018","unstructured":"Darrah E, Giles JT, Davis RL, Naik P, Wang H, Konig MF, et al. Autoantibodies to peptidylarginine deiminase 2 are associated with less severe disease in rheumatoid arthritis. Front Immunol. 2018;9:2696. https:\/\/doi.org\/10.3389\/fimmu.2018.02696.","journal-title":"Front Immunol."},{"issue":"1","key":"2528_CR23","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1186\/s13075-018-1533-z","volume":"20","author":"LC Cappelli","year":"2018","unstructured":"Cappelli LC, Konig MF, Gelber AC, Bingham CO 3rd, Darrah E. Smoking is not linked to the development of anti-peptidylarginine deiminase 4 autoantibodies in rheumatoid arthritis. Arthritis Res Ther. 2018;20(1):59. https:\/\/doi.org\/10.1186\/s13075-018-1533-z.","journal-title":"Arthritis Res Ther."},{"issue":"4","key":"2528_CR24","doi-asserted-by":"publisher","first-page":"516","DOI":"10.1002\/art.40401","volume":"70","author":"MK Demoruelle","year":"2018","unstructured":"Demoruelle MK, Bowers E, Lahey LJ, Sokolove J, Purmalek M, Seto NL, et al. 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 Rheumatol. 2018;70(4):516\u201327. https:\/\/doi.org\/10.1002\/art.40401.","journal-title":"Arthritis Rheumatol."}],"container-title":["Arthritis Research &amp; Therapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13075-021-02528-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13075-021-02528-5\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13075-021-02528-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,6,6]],"date-time":"2021-06-06T22:05:27Z","timestamp":1623017127000},"score":1,"resource":{"primary":{"URL":"https:\/\/arthritis-research.biomedcentral.com\/articles\/10.1186\/s13075-021-02528-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,6]]},"references-count":24,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["2528"],"URL":"https:\/\/doi.org\/10.1186\/s13075-021-02528-5","relation":{},"ISSN":["1478-6362"],"issn-type":[{"value":"1478-6362","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,6]]},"assertion":[{"value":"26 January 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 May 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 June 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All study procedures had ethics approval by the Colorado Multiple Institutional Review Board (#13-2606). Written informed consent was obtained from all subjects.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"ED is an author on licensed patent no. 8,975,033, entitled \u201cHuman autoantibodies specific for pad3 which are cross-reactive with pad4 and their use in the diagnosis and treatment of rheumatoid arthritis and related diseases.\u201d MKD, JMN, VMK, KDD, and ED have received research support from Pfizer Inc. on studies related to RA. ED has received research support from Bristol Myers Squibb and Celgene, and VMH and KDD have research support from Janssen on studies related to RA. All other authors declare that they have no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"163"}}