{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,7]],"date-time":"2026-03-07T14:14:29Z","timestamp":1772892869901,"version":"3.50.1"},"reference-count":28,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T00:00:00Z","timestamp":1772755200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deanship of Scientific Research DSR, Vice-Presidency of Graduated Studies and Scientific Research, King Faisal University","award":["KFU260945"],"award-info":[{"award-number":["KFU260945"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objectives: Epidemiological studies have proven that coxsackievirus B3 (CVB3) is the major virus that causes acute and chronic myocarditis and pancreatitis. Currently, there are no antiviral therapeutic drugs or vaccines that are available for use as clinical therapeutics or vaccines. Subunit polypeptides-based vaccines, especially when combined with adjuvants, represent safe and effective vaccine platforms because they are considered to be better immunogens. The viral capsid protein VP1 of CVB3 is the most immunogenic viral polypeptide, providing opportunities for its use in designing subunit polypeptide vaccines. In the present study, we designed and produced a CVB3 vaccine candidate based on the recombinant expression of the major immunogenic viral protein VP1 of a wild-type CVB3 strain. Methods: We assessed its induced humoral and cellular immune responses and then evaluated its protective immunity against pathogenic CVB3 strain challenges in a Balb\/c mouse model. Neutralizing specific antibodies and cytokine interferon gamma (INF-\u03b3) production were determined in the sera of both prime- and prime-boost-immunized mice with the vaccine candidate. Results: Our results demonstrate that the recombinant rVP1 expressed in a eukaryotic insect cell baculovirus vector system elicited cellular and humoral immune responses, protecting Balb\/c mice from lethal challenges. Conclusions: Hence, the vaccine produced based on the recombinant expression of VP1 is a promising and potential candidate against natural CVB3 infections.<\/jats:p>","DOI":"10.3390\/vaccines14030244","type":"journal-article","created":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T17:25:59Z","timestamp":1772817959000},"page":"244","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Recombinant Viral Capsid Protein rVP1 Induces Protective Immunity Against Coxsackievirus B3 (CVB3) Lethal Challenges in Balb\/c Mouse Model"],"prefix":"10.3390","volume":"14","author":[{"given":"Manel","family":"Ben M\u2019hadheb","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, College of Science, King Faisal University, P.O. Box 380, Al-Ahsa 31982, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1992-7976","authenticated-orcid":false,"given":"Ikbel Hadj","family":"Hassine","sequence":"additional","affiliation":[{"name":"Department of Virology, Health Sciences College, Western University, 1393 Western Road, London, ON N6G 3K7, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0746-488X","authenticated-orcid":false,"given":"Mohammed A.","family":"Almalki","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, College of Science, King Faisal University, P.O. Box 380, Al-Ahsa 31982, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mouna","family":"Hassine","sequence":"additional","affiliation":[{"name":"Virology and Antiviral Strategies Research Unit UR17ES30, College of Biotechnology, Monastir University, BP 74, Tahar Hadded Street, Monastir 5000, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9170-3403","authenticated-orcid":false,"given":"Jawhar","family":"Gharbi","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, College of Science, King Faisal University, P.O. Box 380, Al-Ahsa 31982, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,6]]},"reference":[{"key":"ref_1","first-page":"905","article-title":"The epidemiology and genetics of insulindependent diabetes mellitus","volume":"111","author":"Norris","year":"1987","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1516","DOI":"10.2337\/diacare.23.10.1516","article-title":"Incidence of childhood type 1 diabetes worldwide. Diabetes Mondiale (Dia Mond) project Group","volume":"23","author":"Karvonen","year":"2000","journal-title":"Diabet. Care"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1080\/ann.34.3.138.147","article-title":"Enterovirus infections and type 1 diabetes","volume":"34","year":"2002","journal-title":"Ann. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2835","DOI":"10.1007\/s11033-020-05333-6","article-title":"Characterization of Coxsackievirus B4 virus-like particles VLP produced by the recombinant baculovirus-insect cell system expressing the major capsid protein","volume":"47","author":"Gharbi","year":"2020","journal-title":"Mol. Biol. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1126\/science.1218713","article-title":"Crystal structure of human enterovirus 71","volume":"336","author":"Plevka","year":"2012","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1016\/S0969-2126(01)00201-5","article-title":"The structure of coxsackievirus B3 at 3.5 A resolution","volume":"3","author":"Muckelbauer","year":"1995","journal-title":"Structure"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Gharbi, J., Hadj Hassine, I., Hassine, M., Almalki, M.A., Alyami, A., Albashir, A., and Ben M\u2019hadheb, M. (2023). Viral protein VP1 virus like-particles (VLP) of CVB4 induces protective immunity against lethal challenges with diabetogenic E2 and wild type JBV strains in mice model. Viruses, 15.","DOI":"10.3390\/v15040878"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1038\/337709a0","article-title":"The three-dimensional structure of foot-and-mouth disease virus at 2.9 \u00c5 resolution","volume":"337","author":"Acharya","year":"1989","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1288","DOI":"10.1128\/JCM.37.5.1288-1293.1999","article-title":"Typing of human enteroviruses by partial sequencing of VP1","volume":"37","author":"Oberste","year":"1999","journal-title":"J. Clin. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0166-3542(00)00132-7","article-title":"DNA vaccine-mediated immune responses in Coxsackie virus B3-infected mice","volume":"49","author":"Henke","year":"2001","journal-title":"Antivir. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2944","DOI":"10.1016\/j.vaccine.2005.12.047","article-title":"Oral immunizationof mice using transgenic tomato fruit expressing VP1 protein from enterovirus 71","volume":"24","author":"Chen","year":"2006","journal-title":"Vaccine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1569","DOI":"10.1007\/s10529-014-1523-4","article-title":"Identifying and engineering promoters for high level and sustainable therapeutic recombinant protein production in cultured mammalian cells","volume":"36","author":"Ho","year":"2014","journal-title":"Biotechnol. Lett."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Mandl, J.N., Schneider, C., Schneider, D.S., and Baker, M.L. (2018). Going to bat(s) for studies of disease tolerance. Front. Immunol., 9.","DOI":"10.3389\/fimmu.2018.02112"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/s12951-021-00806-7","article-title":"Virus-like particles: Preparation, immunogenicity and their roles as nanovaccines and drug nanocarriers","volume":"19","author":"Saghi","year":"2021","journal-title":"J. Nanobiotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.4161\/hv.24949","article-title":"Progress on the research and development of inactivated EV71 whole-virus vaccines","volume":"9","author":"Liang","year":"2013","journal-title":"Hum. Vaccine Immunother."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2301","DOI":"10.1016\/j.vaccine.2012.01.061","article-title":"Vaccination with Coxsackievirus B3 virus-like particles elicits humoral immune response and protects mice against myocarditis","volume":"30","author":"Zhang","year":"2012","journal-title":"Vaccine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1186\/s12992-020-00594-z","article-title":"Circulating vaccine-derived poliovirus: A menace to the end game of polio eradication","volume":"16","author":"Ming","year":"2020","journal-title":"Glob. Health"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1128\/CVI.00609-15","article-title":"Brief history and characterization of enhanced respiratory syncytial virus disease","volume":"23","author":"Acosta","year":"2015","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.it.2017.10.002","article-title":"Improving Vaccine and Immunotherapy Design Using Biomaterials","volume":"39","author":"Bookstaver","year":"2018","journal-title":"Trends Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e2402055","DOI":"10.1002\/adhm.202402055","article-title":"Tuning Helical Peptide Nanofibers as a Sublingual Vaccine Platform for a Variety of Peptide Epitopes","volume":"14","author":"Roe","year":"2025","journal-title":"Adv. Healthc. Mater."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method for estimating fifty percent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2564","DOI":"10.3389\/fimmu.2019.02564","article-title":"VP1 of Enterovirus 71 Protects Mice Against enterovirus 71 and Coxsackievirus B3 in Lethal Challenge Experiment","volume":"10","author":"Chen","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_23","unstructured":"CBER (2010). Guidance for Industry, Characterization and Qualification of Cell Substrates and Other Biological Materials Used in the Production of Viral Vaccines for Infectious Disease Indications, Food and Drug Administration (FDA)."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4137","DOI":"10.3389\/fmicb.2021.790121","article-title":"Virus-Like Particles: Revolutionary Platforms for Developing Vaccines Against Emerging Infectious Diseases","volume":"12","author":"Tariq","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s12257-022-0107-8","article-title":"Virus-like Particles as Antiviral Vaccine: Mechanism, Design, and Application","volume":"28","author":"Zhang","year":"2023","journal-title":"Biotechnol. Bioprocess Eng."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Braz Gomes, K., Vijayanand, S., Bagwe, P., Menon, I., Kale, A., Patil, S., Kang, S.-M., Uddin, M.N., and D\u2019Souza, M.J. (2023). Vaccine-Induced Immunity Elicited by Microneedle Delivery of Influenza Ectodomain Matrix Protein 2 Virus-like Particle (M2e VLP)-Loaded PLGA Nanoparticles. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms241310612"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6642","DOI":"10.1016\/j.vaccine.2012.08.071","article-title":"A virus-like particle vaccine for Coxsackievirus A16 potently elicits neutralizing antibodies that protect mice against lethal challenge","volume":"30","author":"Liu","year":"2012","journal-title":"Vaccine"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Xing, J., Zhao, X., Li, X., Fang, R., Sun, M., Zhang, Y., and Song, N. (2025). The recent advances in vaccine adjuvants. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1557415"}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/3\/244\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T17:32:57Z","timestamp":1772818377000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/3\/244"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,6]]},"references-count":28,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["vaccines14030244"],"URL":"https:\/\/doi.org\/10.3390\/vaccines14030244","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,6]]}}}