{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:41:08Z","timestamp":1763642468676,"version":"3.37.3"},"reference-count":18,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2018,10,24]],"date-time":"2018-10-24T00:00:00Z","timestamp":1540339200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["16K10141 to HI"],"award-info":[{"award-number":["16K10141 to HI"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Oral Pathology Medicine"],"published-print":{"date-parts":[[2019,1]]},"abstract":"Background<\/jats:title>Mucous membrane pemphigoid (MMP<\/jats:styled-content>) is a rare chronic autoimmune subepithelial blistering disorder, targeting multiple basement membrane zone (BMZ<\/jats:styled-content>) proteins including collagen XVII<\/jats:styled-content> (COL<\/jats:styled-content>17). Circulating autoantibodies of MMP<\/jats:styled-content> are often undetected due to their lower titers. The oral mucosa is a valuable substrate for the detection of autoantibodies in MMP<\/jats:styled-content> patients. However, obtaining normal human oral mucosa is more difficult than obtaining normal human skin. We established immortalized normal human oral mucosal keratinocytes (OMK<\/jats:styled-content>s) and performed immunoblotting using immortalized OMK<\/jats:styled-content> lysate for detecting autoantigens in MMP<\/jats:styled-content>.<\/jats:p><\/jats:sec>Methods<\/jats:title>Immortalized OMK<\/jats:styled-content>s were generated from primary OMK<\/jats:styled-content>s using E6\/E7 proteins of HPV<\/jats:styled-content>. We compared the protein expression levels of major BMZ<\/jats:styled-content> proteins between primary OMK<\/jats:styled-content>s and immortalized OMK<\/jats:styled-content>s. We performed immunoblotting to detect autoantigens using cell lysates from immortalized OMK<\/jats:styled-content>s in 30 MMP<\/jats:styled-content> patients.<\/jats:p><\/jats:sec>Results<\/jats:title>There were no significant differences between primary OMK<\/jats:styled-content>s and immortalized OMK<\/jats:styled-content>s in terms of protein expression levels of the BMZ<\/jats:styled-content> proteins, including COL<\/jats:styled-content>17, laminin 332, integrin \u03b16\/\u03b24, collagen VII<\/jats:styled-content>, and collagen IV<\/jats:styled-content>. Cell lysates of immortalized OMK<\/jats:styled-content>s effectively identified MMP<\/jats:styled-content> autoantigens in 60% (18\/30) of MMP<\/jats:styled-content> sera. We found an interesting case of MMP<\/jats:styled-content> whose autoantibodies preferentially reacted to the 120\u2010kD<\/jats:styled-content> protein that is an ectodomain of COL<\/jats:styled-content>17.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>We demonstrated that a cell lysate of immortalized OMK<\/jats:styled-content>s is a reliable substrate for the detection of MMP<\/jats:styled-content> autoantigens. This newly developed immunoblotting analysis method promises to contribute to the diagnosis of MMP<\/jats:styled-content>.<\/jats:p><\/jats:sec>","DOI":"10.1111\/jop.12780","type":"journal-article","created":{"date-parts":[[2018,9,17]],"date-time":"2018-09-17T14:14:59Z","timestamp":1537193699000},"page":"60-67","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["The identification of autoantigens in mucous membrane pemphigoid using immortalized oral mucosal keratinocytes"],"prefix":"10.1111","volume":"48","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4055-9047","authenticated-orcid":false,"given":"Mayumi","family":"Kamaguchi","sequence":"first","affiliation":[{"name":"Department of Oral Diagnosis and Medicine Hokkaido University Graduate School of Dental Medicine Sapporo Japan"},{"name":"Department of Dermatology Hokkaido University Graduate School of Medicine Sapporo 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