{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T01:25:08Z","timestamp":1747185908123,"version":"3.40.5"},"reference-count":25,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2021,12,8]],"date-time":"2021-12-08T00:00:00Z","timestamp":1638921600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Diabetes Invest"],"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Aims\/Introduction<\/jats:title><jats:p>The need for antiserum for immunohistochemical (IHC) detection of enterovirus (EV) in formaldehyde\u2010fixed and paraffin\u2010embedded samples is increasing. The gold standard monoclonal antibody (clone 5D8\/1) against EV\u2010envelope protein (VP1) was proven to cross\u2010react with other proteins. Another candidate marker of EV proteins is 2A\u2009protease (2A<jats:sup>pro<\/jats:sup>), which is encoded by the EV gene and translated by the host cells during EV replication, and participates processing proproteins to viral capsid proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and Methods<\/jats:title><jats:p>We raised polyclonal antiserum by immunizing a rabbit with an 18\u2010mer peptide of Coxsackievirus\u2009B1 (CVB1)\u20102A<jats:sup>pro<\/jats:sup>, and examined the specificity and sensitivity for EV on formaldehyde\u2010fixed and paraffin\u2010embedded tissue samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Enzyme\u2010linked immunosorbent assay study showed a high titer of antibody for 18\u2010mer peptide of CVB1\u20102A<jats:sup>pro<\/jats:sup>, cross\u2010reacting with CVB3\u20102A<jats:sup>pro<\/jats:sup> peptide. IHC showed that antiserum against 2A<jats:sup>pro<\/jats:sup> reacted with CVB1\u2010infected and VP1\u2010positive Vero cells. Confocal laser scanning microscopy showed that antigen stained by the 2A<jats:sup>pro<\/jats:sup> antibody located in the same cell with VP1 stained by 5D8\/1. IHC using 2A<jats:sup>pro<\/jats:sup> antiserum showed dense staining in the islets of EV\u2010associated fulminant type\u20091 diabetes pancreas and that located in the same cell stained positive for VP1 (5D8\/1). Specificity of 2A<jats:sup>pro<\/jats:sup> antiserum by IHC staining was confirmed by negative 2A<jats:sup>pro<\/jats:sup> in 14 VP1\u2010negative non\u2010diabetes control pancreases.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our study provides a new polyclonal antiserum against CVB1\u20102A<jats:sup>pro<\/jats:sup>, which might be useful for IHC of EV\u2010infected human tissues stored as archive of formaldehyde\u2010fixed and paraffin\u2010embedded tissue samples.<\/jats:p><\/jats:sec>","DOI":"10.1111\/jdi.13700","type":"journal-article","created":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T22:57:32Z","timestamp":1634770652000},"page":"435-442","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Immunohistochemical detection of enteroviruses in pancreatic tissues of patients with type\u20091 diabetes using a polyclonal antibody against 2A\u2009protease of Coxsackievirus"],"prefix":"10.1111","volume":"13","author":[{"given":"Erika","family":"Jimbo","sequence":"first","affiliation":[{"name":"Division of Immunology and Molecular Medicine Okinaka Memorial Institute for Medical Research Tokyo Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4364-2527","authenticated-orcid":false,"given":"Tetsuro","family":"Kobayashi","sequence":"additional","affiliation":[{"name":"Division of Immunology and Molecular Medicine Okinaka Memorial Institute for Medical Research Tokyo Japan"},{"name":"Department of Endocrinology and Metabolism Toranomon Hospital Tokyo Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akira","family":"Takeshita","sequence":"additional","affiliation":[{"name":"Department of Endocrinology and Metabolism Toranomon Hospital Tokyo 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