{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T17:25:40Z","timestamp":1761845140239,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,3,21]],"date-time":"2020-03-21T00:00:00Z","timestamp":1584748800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"publisher","award":["18TPA34170206"],"award-info":[{"award-number":["18TPA34170206"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Coxsackievirus group B (CVB) contains six serotypes that can affect various organs. Some of these organ-specific diseases such as myocarditis and pancreatitis can be caused by more than one serotype. Thus, development of immunological tools common to multiple serotypes is desired. This is especially critical for analyzing antigen-specific T cell responses at a single cell level. To this end, we made efforts to identify the immunogenic epitopes of CVB3 leading us to localize three T cell epitopes within the viral protein 1 (VP1) namely, VP1 681\u2013700, VP1 721\u2013740 and VP1 771\u2013790. First, we confirmed their immunogenicity in the immunization settings. Second, we sought to verify the ability of VP1 epitopes to bind major histocompatibility complex (MHC) class II (IAk) molecules. Third, we created MHC class II (IAk) dextramers and tetramers and ascertained the T cell responses to be antigen-specific. Fourth, we analyzed the T cell responses in animals infected with CVB3 and noted the magnitude of antigen-specific T cell responses occurring in the order of VP1 721\u2013740 and VP1 681\u2013700 followed by VP1 771\u2013790 as verified by proliferation assay and IAk tetramer staining. All epitopes induced interferon (IFN)-\u03b3 as a major cytokine. Finally, we investigated whether the VP1 tools generated for CVB3 can also be used to verify T cell responses in infections caused by other serotypes. To this end, we established the CVB4 infection model in A\/J mice and found that the CVB4 infection led to the induction of IFN-\u03b3-producing T cell responses primarily for VP1 721\u2013740 and VP1 681\u2013700. Thus, the VP1-specific tools, particularly IAk tetramers can be used to monitor anti-viral T cell responses in multiple CVB serotypes.<\/jats:p>","DOI":"10.3390\/v12030347","type":"journal-article","created":{"date-parts":[[2020,3,23]],"date-time":"2020-03-23T04:03:01Z","timestamp":1584936181000},"page":"347","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Identification of Immunogenic Epitopes That Permit the Detection of Antigen-Specific T Cell Responses in Multiple Serotypes of Group B Coxsackievirus Infections"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6762-2654","authenticated-orcid":false,"given":"Ninaad","family":"Lasrado","sequence":"first","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6307-9391","authenticated-orcid":false,"given":"Arunakumar","family":"Gangaplara","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajkumar","family":"Arumugam","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chandirasegaran","family":"Massilamany","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sayli","family":"Pokal","sequence":"additional","affiliation":[{"name":"Department of Statistics, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuzhen","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Statistics, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shi-Hua","family":"Xiang","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5166-2296","authenticated-orcid":false,"given":"David","family":"Steffen","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4082-9254","authenticated-orcid":false,"given":"Jay","family":"Reddy","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68503, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1542\/pir.19.6.183","article-title":"Enterovirus infections","volume":"19","author":"Zaoutis","year":"1998","journal-title":"Pediatr. Rev. Am. Acad. Pediatr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1016\/j.virol.2010.12.014","article-title":"Enterovirus infections of the central nervous system","volume":"411","author":"Rhoades","year":"2011","journal-title":"Virology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/1096-9071(200009)62:1<70::AID-JMV11>3.0.CO;2-R","article-title":"Group B coxsackievirus myocarditis and pancreatitis: Connection between viral virulence phenotypes in mice","volume":"62","author":"Tracy","year":"2000","journal-title":"J. Med. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1093\/eurheartj\/12.suppl_D.56","article-title":"Molecular probes for detection of persisting enterovirus infection of human heart and their prognostic value","volume":"12","author":"Archard","year":"1991","journal-title":"Eur. Heart J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/S0065-2776(08)00604-4","article-title":"Pathogenesis of myocarditis and dilated cardiomyopathy","volume":"99","author":"Cihakova","year":"2008","journal-title":"Adv. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1161\/01.CIR.0000155616.07901.35","article-title":"High prevalence of viral genomes and multiple viral infections in the myocardium of adults with \u201cidiopathic\u201d left ventricular dysfunction","volume":"111","author":"Kuhl","year":"2005","journal-title":"Circulation"},{"key":"ref_7","unstructured":"Martino, T., Liu, P., and Sole, M.J. (1995). Enteroviral myocarditis and dialted cardiomyopathy: A review of clinical and experimental studies. Human Enterovirus Infections, ASM."},{"key":"ref_8","first-page":"275","article-title":"Persistent coxsackievirus infection: Enterovirus persistence in chronic myocarditis and dilated cardiomyopathy","volume":"323","author":"Chapman","year":"2008","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1111\/j.1749-6632.1986.tb20864.x","article-title":"Postinfectious autoimmunity: Two distinct phases of coxsackievirus B3-induced myocarditis","volume":"475","author":"Rose","year":"1986","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1056\/NEJM197905243002102","article-title":"Isolation of a virus from the pancreas of a child with diabetic ketoacidosis","volume":"300","author":"Yoon","year":"1979","journal-title":"N. Engl. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5115","DOI":"10.1073\/pnas.0700442104","article-title":"Coxsackie B4 virus infection of beta cells and natural killer cell insulitis in recent-onset type 1 diabetic patients","volume":"104","author":"Dotta","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/S0140-6736(95)91270-3","article-title":"Coxsackie B virus infection and onset of childhood diabetes","volume":"346","author":"Clements","year":"1995","journal-title":"Lancet"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1136\/bmj.3.5671.627","article-title":"Viral antibodies in diabetes mellitus","volume":"3","author":"Gamble","year":"1969","journal-title":"Br. Med. J."},{"key":"ref_14","first-page":"259","article-title":"The CVB and etiology of type 1 diabetes","volume":"323","author":"Drescher","year":"2008","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1196\/annals.1394.009","article-title":"Coxsackievirus infections and NOD mice: Relevant models of protection from, and induction of, type 1 diabetes","volume":"1103","author":"Tracy","year":"2007","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"12097","DOI":"10.1128\/JVI.76.23.12097-12111.2002","article-title":"Toward testing the hypothesis that group B coxsackieviruses (CVB) trigger insulin-dependent diabetes: Inoculating nonobese diabetic mice with CVB markedly lowers diabetes incidence","volume":"76","author":"Tracy","year":"2002","journal-title":"J. Virol."},{"key":"ref_17","first-page":"1515","article-title":"Immunoregulatory mechanisms triggered by viral infections protect from type 1 diabetes in mice","volume":"119","author":"Filippi","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.2353\/ajpath.2008.070207","article-title":"Interleukin-13 protects against experimental autoimmune myocarditis by regulating macrophage differentiation","volume":"172","author":"Cihakova","year":"2008","journal-title":"Am. J. Pathol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.ijcard.2015.09.016","article-title":"Localization of CD8 T cell epitope within cardiac myosin heavy chain-\u03b1 334\u2013352 that induces autoimmune myocarditis in A\/J mice","volume":"202","author":"Massilamany","year":"2016","journal-title":"Int. J. Cardiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3160","DOI":"10.1016\/j.ajpath.2016.08.005","article-title":"Identification of an Epitope from Adenine Nucleotide Translocator 1 That Induces Inflammation in Heart in A\/J Mice","volume":"186","author":"Basavalingappa","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1002\/iid3.177","article-title":"Branched chain \u03b1-ketoacid dehydrogenase kinase 111\u2013130, a T cell epitope that induces both autoimmune myocarditis and hepatitis in A\/J mice","volume":"5","author":"Krishnan","year":"2017","journal-title":"Immun. Inflamm. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.cellimm.2011.08.013","article-title":"Identification of novel mimicry epitopes for cardiac myosin heavy chain-\u03b1 that induce autoimmune myocarditis in A\/J mice","volume":"271","author":"Massilamany","year":"2011","journal-title":"Cell. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.jim.2011.07.004","article-title":"Detection of cardiac myosin heavy chain-alpha-specific CD4 cells by using MHC class II\/IA(k) tetramers in A\/J mice","volume":"372","author":"Massilamany","year":"2011","journal-title":"J. Immunol. Methods"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"870","DOI":"10.4049\/jimmunol.170.2.870","article-title":"Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers","volume":"170","author":"Reddy","year":"2003","journal-title":"J. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"831","DOI":"10.1172\/JCI200318509","article-title":"Ex vivo analysis of human memory CD4 T cells specific for hepatitis C virus using MHC class II tetramers","volume":"112","author":"Day","year":"2003","journal-title":"J. Clin. Investig."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1723","DOI":"10.1084\/jem.189.11.1723","article-title":"pH-dependent peptide binding properties of the type I diabetes-associated I-Ag7 molecule: Rapid release of CLIP at an endosomal pH","volume":"189","author":"Hausmann","year":"1999","journal-title":"J. Exp. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1007\/BF03401615","article-title":"Quantitative analysis of peptides from myelin basic protein binding to the MHC class II protein, I-Au, which confers susceptibility to experimental allergic encephalomyelitis","volume":"2","author":"Fugger","year":"1996","journal-title":"Mol. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4175","DOI":"10.4049\/jimmunol.171.8.4175","article-title":"Ex vivo analysis of thymic CD4 T cells in nonobese diabetic mice with tetramers generated from I-A(g7)\/class II-associated invariant chain peptide precursors","volume":"171","author":"Jang","year":"2003","journal-title":"J. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Massilamany, C., Upadhyaya, B., Gangaplara, A., Kuszynski, C., and Reddy, J. (2011). Detection of autoreactive CD4 T cells using major histocompatibility complex class II dextramers. BMC Immunol., 12.","DOI":"10.1186\/1471-2172-12-40"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.jneuroim.2009.11.006","article-title":"An epitope from Acanthamoeba castellanii that cross-react with proteolipid protein 139-151-reactive T cells induces autoimmune encephalomyelitis in SJL mice","volume":"219","author":"Massilamany","year":"2010","journal-title":"J. Neuroimmunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.jneuroim.2010.09.011","article-title":"Gender differences in CNS autoimmunity induced by mimicry epitope for PLP 139-151 in SJL mice","volume":"230","author":"Massilamany","year":"2011","journal-title":"J. Neuroimmunol."},{"key":"ref_32","unstructured":"Daniel, W.W. (1978). Applied Nonparametric Statistics, Houghton Mifflin."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1038\/156177a0","article-title":"A New Test for 2 \u00d7 2 Tables","volume":"156","author":"Barnard","year":"1945","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4508","DOI":"10.4049\/jimmunol.153.10.4508","article-title":"A predominant viral epitope recognized by T cells from the periphery and demyelinating lesions of SJL\/J mice infected with Theiler\u2019s virus is located within VP1(233-244)","volume":"153","author":"Yauch","year":"1994","journal-title":"J. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2037","DOI":"10.1084\/jem.170.6.2037","article-title":"Alteration of amino acid 101 within capsid protein VP-1 changes the pathogenicity of Theiler\u2019s murine encephalomyelitis virus","volume":"170","author":"Zurbriggen","year":"1989","journal-title":"J. Exp. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/0306-9877(88)90041-2","article-title":"Sequence homology between encephalomyocarditis virus protein VPI and histidyl-tRNA synthetase supports a hypothesis of molecular mimicry in polymyositis","volume":"25","author":"Walker","year":"1988","journal-title":"Med. Hypotheses"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1038\/317359a0","article-title":"Binding of immunogenic peptides to Ia histocompatibility molecules","volume":"317","author":"Babbitt","year":"1985","journal-title":"Nature"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1567","DOI":"10.3389\/fimmu.2017.01567","article-title":"beta1-Adrenergic Receptor Contains Multiple IA(k) and IE(k) Binding Epitopes That Induce T Cell Responses with Varying Degrees of Autoimmune Myocarditis in A\/J Mice","volume":"8","author":"Basavalingappa","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Massilamany, C., Gangaplara, A., Jia, T., Elowsky, C., Kang, G., Riethoven, J.-J., Li, Q., Zhou, Y., and Reddy, J. (2014). Direct staining with major histocompatibility complex class II dextramers permits detection of antigen-specific, autoreactive CD4 T cells in situ. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0087519"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1107\/S0907444995002253","article-title":"Structure determination of coxsackievirus B3 to 3.5 A resolution","volume":"51","author":"Muckelbauer","year":"1995","journal-title":"Acta Cryst. D Biol. Cryst."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/j.diabet.2008.05.008","article-title":"Role of coxsackievirus B4 in the pathogenesis of type 1 diabetes","volume":"34","author":"Jaidane","year":"2008","journal-title":"Diabetes Metab."},{"key":"ref_42","first-page":"67","article-title":"T lymphocyte responses in CVB3-induced murine myocarditis","volume":"88","author":"Huber","year":"1993","journal-title":"Scand. J. Infect. Dis. Suppl."},{"key":"ref_43","first-page":"899","article-title":"Coxsackievirus-induced disease. CD4+ cells initiate both myocarditis and pancreatitis in DBA\/2 mice","volume":"135","author":"Blay","year":"1989","journal-title":"Am. J. Pathol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.1099\/0022-1317-73-6-1387","article-title":"Coxsackievirus B4 infection of the mouse pancreas: Acute and persistent infection","volume":"73","author":"Vella","year":"1992","journal-title":"J. Gen. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1128\/JVI.80.3.1451-1462.2006","article-title":"Structure and dynamics of coxsackievirus B4 2A proteinase, an enyzme involved in the etiology of heart disease","volume":"80","author":"Baxter","year":"2006","journal-title":"J. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1097\/BOR.0b013e328353372d","article-title":"Update on coxsackievirus B3 myocarditis","volume":"24","author":"Fairweather","year":"2012","journal-title":"Curr. Opin. Rheumatol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1111\/j.1365-2141.2000.02391.x","article-title":"The reconstitution of CD45RBhiCD4+ naive T cells is inversely correlated with donor age in murine allogeneic haematopoietic stem cell transplantation","volume":"111","author":"Hirayama","year":"2000","journal-title":"Br. J. Haematol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1505","DOI":"10.1084\/jem.187.9.1505","article-title":"Evidence that the autoimmune antigen myelin basic protein (MBP) Ac1-9 binds towards one end of the major histocompatibility complex (MHC) cleft","volume":"187","author":"Lee","year":"1998","journal-title":"J. Exp. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"936","DOI":"10.1002\/eji.201040466","article-title":"Bystander activation of CD4+ T cells","volume":"40","author":"Boyman","year":"2010","journal-title":"Eur. J. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1745","DOI":"10.2337\/diabetes.51.6.1745","article-title":"Characterization of the T-cell response to coxsackievirus B4: Evidence that effector memory cells predominate in patients with type 1 diabetes","volume":"51","author":"Ellis","year":"2002","journal-title":"Diabetes"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"D339","DOI":"10.1093\/nar\/gky1006","article-title":"The Immune Epitope Database (IEDB): 2018 update","volume":"47","author":"Vita","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1016\/j.virol.2015.06.006","article-title":"Recent progress in understanding coxsackievirus replication, dissemination, and pathogenesis","volume":"484","author":"Sin","year":"2015","journal-title":"Virology"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/3\/347\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:10:27Z","timestamp":1760173827000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/3\/347"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,21]]},"references-count":52,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2020,3]]}},"alternative-id":["v12030347"],"URL":"https:\/\/doi.org\/10.3390\/v12030347","relation":{},"ISSN":["1999-4915"],"issn-type":[{"type":"electronic","value":"1999-4915"}],"subject":[],"published":{"date-parts":[[2020,3,21]]}}}