{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T23:05:30Z","timestamp":1770073530212,"version":"3.49.0"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T00:00:00Z","timestamp":1734998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015103","name":"Japan Racing Association","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100015103","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"AMED","doi-asserted-by":"publisher","award":["JP21zf0127001"],"award-info":[{"award-number":["JP21zf0127001"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The development of a hepatitis E virus (HEV) vaccine is critical, with ORF2 capsid protein as the main target. We previously demonstrated that oral coadministration of recombinant ORF2 with immunomodulatory bacterium-like-particles (IBLP) induces a specific immune response in mice, particularly using IBLP derived from <jats:italic>Lacticaseibacillus rhamnosus<\/jats:italic> IBL027 (IBLP027), which was effective in eliciting a local humoral response. IBLP are non-live bacteria with adjuvant and carrier properties, serving as a platform for exposing proteins or antigens fused to LysM (lysine motif) domains, protein modules that bind to cell wall polysaccharides like peptidoglycan.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials<\/jats:title><jats:p>We cloned the most immunogenic domain of ORF2 (O2P2) fused to five LysM domains (LysM<jats:sub>5<\/jats:sub>O2P2) and displayed this chimeric protein on the surface of IBLP027 to create a prototype vaccine (IBLP027-LysM<jats:sub>5<\/jats:sub>O2P2). We evaluated its capacity to induce an immune response <jats:italic>in vivo<\/jats:italic> by immunizing mice with three doses of either the experimental vaccine or the chimeric protein alone, using an oral or a combined schedule with subcutaneous priming followed by oral boosting. Control groups received IBLP027. Sera and small intestine fluid were analyzed for humoral response, while Peyer\u2019s patches and spleen immune cells were used for ex vivo stimulation with capsid protein to assess cellular response.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The oral scheme failed to elicit an IgG response, but this was overcome by a subcutaneous priming dose followed by oral boosters, which led to increasing IgG titers in the combined scheme. The highest IgG titers were seen in the vaccine prototype group. Most groups produced significantly higher IgA levels in intestinal fluid, especially in those that received the oral scheme. Cellular response studies showed increased tumor necrosis factor (TNF)-\u03b1, interferon (IFN)-\u03b3 interleukin (IL)-4, and IL-17 levels in groups receiving the chimeric protein via oral or combined schedules.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Further and continuous research is needed to better understand both the needs and expectations of students and supervisors in different academic realities, including in Veterinary Medicine schools, from which the information available on the subject is scarce.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2024.1512018","type":"journal-article","created":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T09:18:08Z","timestamp":1735031888000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["An experimental chimeric hepatitis E virus vaccine elicits both local and systemic immune responses"],"prefix":"10.3389","volume":"15","author":[{"given":"Melisa Florencia","family":"M\u00fcller","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacinto","family":"Sacur","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia Matias","family":"Brancher","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Daniela","family":"Vera","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorena","family":"Arce","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Fernanda","family":"Raya-Tonetti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haruki","family":"Kitazawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julio","family":"Villena","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda Guadalupe","family":"Vizoso-Pinto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,12,24]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"961","DOI":"10.1007\/s12602-019-09598-7","article-title":"Oral vaccination with hepatitis E virus capsid protein and immunobiotic bacterium-like particles induce intestinal and systemic immunity in mice","volume":"12","author":"Arce","year":"2020","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1099\/jmm.0.05472-0","article-title":"An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci","volume":"53","author":"Berm\u00fadez-Humar\u00e1n","year":"2004","journal-title":"J. Med. Microbiol."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"880","DOI":"10.1128\/AEM.72.1.880-889.2006","article-title":"Novel surface display system for proteins on non-genetically modified gram-positive bacteria","volume":"72","author":"Bosma","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"517","DOI":"10.1093\/infdis\/170.3.517","article-title":"Epidemic hepatitis E in Pakistan: patterns of serologic response and evidence that antibody to hepatitis E virus protects against disease","volume":"170","author":"Bryan","year":"1994","journal-title":"J. Infect. Dis."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"2457","DOI":"10.1007\/s00705-022-05575-8","article-title":"Structural aspects of hepatitis E virus","volume":"167","author":"Cancela","year":"2022","journal-title":"Arch. Virol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1025","DOI":"10.1056\/NEJMc0903778","article-title":"Persistent carriage of hepatitis E virus in patients with HIV infection","volume":"361","author":"Dalton","year":"2009","journal-title":"N. Engl. J. Med."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1111\/j.1574-6968.2006.00122.x","article-title":"Protein cell surface display in gram-positive bacteria: from single protein to macromolecular protein structure","volume":"256","author":"Desvaux","year":"2006","journal-title":"FEMS Microbiol. Lett."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"905","DOI":"10.1016\/j.clinre.2020.01.010","article-title":"Adjuvanted recombinant HBV vaccine (HBV-AS04) is effective over extended follow-up in dialysis population. An open-label non randomized trial","volume":"44","author":"Fabrizi","year":"2020","journal-title":"Clin. Res. Hepatol. Gastroenterol."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1016\/j.intimp.2014.10.032","article-title":"Oral immunization with recombinant hepatitis E virus antigen displayed on the Lactococcus lactis surface enhances ORF2-specific mucosal and systemic immune responses in mice","volume":"24","author":"Gao","year":"2015","journal-title":"Int. Immunopharmacol."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"e13560","DOI":"10.1371\/journal.pone.0013560","article-title":"Profile of acute infectious markers in sporadic hepatitis E","volume":"5","author":"Huang","year":"2010","journal-title":"PLoS One"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1016\/S0140-6736(23)02234-1","article-title":"Long-term efficacy of a recombinant hepatitis E vaccine in adults: 10-year results from a randomised, double-blind, placebo-controlled, phase 3 trial","volume":"403","author":"Huang","year":"2024","journal-title":"Lancet"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1093\/infdis\/jiae148","article-title":"A phase 1, double-blinded, placebo-controlled clinical trial to evaluate the safety and immunogenicity of HEV-239 (Hecolin\u00ae) vaccine in healthy US adults","volume":"230","author":"Kao","year":"2024","journal-title":"J. Infect. Dis."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"767","DOI":"10.1093\/intimm\/dxab056","article-title":"Mucosal vaccines: wisdom from now and then","volume":"33","author":"Kiyono","year":"2021","journal-title":"Int. Immunol."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1038\/s41541-024-00806-2","article-title":"Three immunizations with Novavax\u2019s protein vaccines increase antibody breadth and provide durable protection from SARS-CoV-2","volume":"9","author":"Lenart","year":"2024","journal-title":"npj Vaccines"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"799","DOI":"10.3390\/v11090799","article-title":"A novel bacterium-like particle vaccine displaying the MERS-CoV receptor-binding domain induces specific mucosal and systemic immune responses in mice","volume":"11","author":"Li","year":"2019","journal-title":"Viruses"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1007\/s00281-009-0191-2","article-title":"Th17 cytokines and vaccine-induced immunity","volume":"32","author":"Lin","year":"2010","journal-title":"Semin. Immunopathol."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"e12426","DOI":"10.1111\/hel.12426","article-title":"Nongenetically modified Lactococcus lactis-adjuvanted vaccination enhanced innate immunity against Helicobacter pylori","volume":"22","author":"Liu","year":"2017","journal-title":"Helicobacter"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"232","DOI":"10.1016\/j.jinf.2019.01.004","article-title":"Hepatitis E virus genotype 3 and capsid protein in the blood and urine of immunocompromised patients","volume":"78","author":"Marion","year":"2019","journal-title":"J. Infect."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"5633","DOI":"10.4049\/jimmunol.179.9.5633","article-title":"Perspectives on mucosal vaccines: is mucosal tolerance a barrier?","volume":"179","author":"Mestecky","year":"2007","journal-title":"J. Immunol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1186\/s12934-016-0468-9","article-title":"Display of recombinant proteins at the surface of lactic acid bacteria: strategies and applications","volume":"15","author":"Michon","year":"2016","journal-title":"Microb. Cell Fact."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"e0011661","DOI":"10.1371\/journal.pntd.0011661","article-title":"Vaccination coverage and adverse events following a reactive vaccination campaign against hepatitis E in Bentiu displaced persons camp, South Sudan","volume":"18","author":"Nesbitt","year":"2024","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"1059","DOI":"10.1016\/j.vaccine.2022.12.064","article-title":"Immunogenicity and safety of a two-dose regimen with hepatitis E virus vaccine in healthy adults in rural Bangladesh: a randomized, double-blind, controlled, phase 2\/pilot trial","volume":"41","author":"\u00d8verb\u00f8","year":"2023","journal-title":"Vaccine"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1310","DOI":"10.1016\/S1473-3099(16)30371-1","article-title":"Unravelling mucosal immunity to poliovirus","volume":"16","author":"Parker","year":"2016","journal-title":"Lancet Infect. Dis."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"869","DOI":"10.3201\/eid1805.111712","article-title":"Hepatitis E virus infection among solid organ transplant recipients, the Netherlands","volume":"18","author":"Pas","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"1098","DOI":"10.1016\/j.vaccine.2022.01.025","article-title":"Oral subunit SARS-CoV-2 vaccine induces systemic neutralizing IgG, IgA and cellular immune responses and can boost neutralizing antibody responses primed by an injected vaccine","volume":"40","author":"Pitcovski","year":"2022","journal-title":"Vaccine"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"15","DOI":"10.3389\/fimmu.2020.00015","article-title":"Immunomodulatory properties of bacterium-like particles obtained from Immunobiotic lactobacilli: prospects for their use as mucosal adjuvants","volume":"11","author":"Raya Tonetti","year":"2020","journal-title":"Front. Immunol."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"21691","DOI":"10.1038\/s41598-021-01087-8","article-title":"Novel LysM motifs for antigen display on lactobacilli for mucosal immunization","volume":"11","author":"Raya-Tonetti","year":"2021","journal-title":"Sci. Rep."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"e2086","DOI":"10.1002\/rmv.2086","article-title":"Updates in hepatitis E virus (HEV) field; lessons learned from human liver chimeric mice","volume":"30","author":"Sayed","year":"2020","journal-title":"Rev. Med. Virol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"1067","DOI":"10.1111\/jvh.12730","article-title":"Risk factors for vertical transmission of hepatitis E virus infection","volume":"24","author":"Sharma","year":"2017","journal-title":"J. Viral Hepat."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"355","DOI":"10.1128\/CVI.00354-10","article-title":"Mucosal IgA responses in healthy adult volunteers following intranasal spray delivery of a live attenuated measles vaccine","volume":"18","author":"Simon","year":"2011","journal-title":"Clin. Vaccine Immunol."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"598","DOI":"10.3389\/fimmu.2018.00598","article-title":"Distinct immunologic properties of soluble versus particulate antigens","volume":"9","author":"Snapper","year":"2018","journal-title":"Front. Immunol."},{"key":"ref32","article-title":"Hepatitis E vaccines background paper Strategic Advisory Group of Experts (SAGE) on Immunization meeting","year":"2024"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"1082273","DOI":"10.3389\/fimmu.2023.1082273","article-title":"The adjuvant effect of bacterium-like particles depends on the route of administration","volume":"14","author":"Sudo","year":"2023","journal-title":"Front. Immunol."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1016\/j.immuni.2007.08.013","article-title":"Stimulation of the intracellular bacterial sensor NOD2 programs dendritic cells to promote interleukin-17 production in human memory T cells","volume":"27","author":"van Beelen","year":"2007","journal-title":"Immunity"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"699","DOI":"10.3389\/fphys.2020.00699","article-title":"The modulation of mucosal antiviral immunity by immunobiotics: could they offer any benefit in the SARS-CoV-2 pandemic?","volume":"11","author":"Villena","year":"2020","journal-title":"Front. Physiol."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1139\/W08-077","article-title":"Oral immunization with recombinant Lactococcus lactis confers protection against respiratory pneumococcal infection","volume":"54","author":"Villena","year":"2008","journal-title":"Can. J. Microbiol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"3747","DOI":"10.1016\/j.vaccine.2018.06.038","article-title":"Development and validation of a monocyte activation test for the control\/safety testing of an OMV-based meningococcal B vaccine","volume":"37","author":"Vipond","year":"2019","journal-title":"Vaccine"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1186\/1743-422X-7-165","article-title":"A systematic approach for the identification of novel, serologically reactive recombinant Varicella-Zoster Virus (VZV) antigens","volume":"7","author":"Vizoso Pinto","year":"2010","journal-title":"Virol. J."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"5988","DOI":"10.1128\/JVI.79.10.5988-5995.2005","article-title":"Virus-specific CD4+ and CD8+ cytotoxic T-cell responses and long-term T-cell memory in individuals vaccinated against polio","volume":"79","author":"Wahid","year":"2005","journal-title":"J. Virol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"a033472","DOI":"10.1101\/cshperspect.a033472","article-title":"Adaptive immune responses in hepatitis a virus and hepatitis E virus infections","volume":"9","author":"Walker","year":"2019","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"2022","DOI":"10.1111\/1751-7915.14022","article-title":"Vaccine with bacterium-like particles displaying HIV-1 gp120 trimer elicits specific mucosal responses and neutralizing antibodies in rhesus macaques","volume":"15","author":"Wang","year":"2022","journal-title":"Microb. Biotechnol."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"e1961","DOI":"10.1002\/rmv.1961","article-title":"An overview: rabbit hepatitis E virus (HEV) and rabbit providing an animal model for HEV study","volume":"28","author":"Wang","year":"2018","journal-title":"Rev. Med. Virol."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"204993611983716","DOI":"10.1177\/2049936119837162","article-title":"Hepatitis E: an underestimated emerging threat","volume":"6","author":"Webb","year":"2019","journal-title":"Ther. Adv. Infect. Dis."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"1738","DOI":"10.1038\/s41598-018-20137-2","article-title":"Dimerization: a structural feature for the protection of hepatitis E virus capsid protein against trypsinization","volume":"8","author":"Wei","year":"2018","journal-title":"Sci. Rep."},{"key":"ref45","article-title":"Hepatitis E vaccine: WHO position paper, May 2015","year":"2015"},{"key":"ref46","article-title":"Hepatitis E: fact sheet","year":"2023"},{"key":"ref47","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1186\/s12985-020-01343-9","article-title":"Hepatitis E virus infection during pregnancy","volume":"17","author":"Wu","year":"2020","journal-title":"Virol. J."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"883","DOI":"10.3390\/v11100883","article-title":"Hepatitis E virus entry","volume":"11","author":"Yin","year":"2019","journal-title":"Viruses"},{"key":"ref49","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1271\/bbb.66.963","article-title":"Antigen presentation by Peyer\u2019s patch cells can induce both Th1-and Th2-type responses depending on antigen dosage, but a different cytokine response pattern from that of spleen cells","volume":"66","author":"Yoshida","year":"2002","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref50","doi-asserted-by":"publisher","first-page":"2290","DOI":"10.1016\/j.vaccine.2024.02.077","article-title":"Safety and immunogenicity of Innovax bivalent human papillomavirus vaccine in girls 9-14 years of age: interim analysis from a phase 3 clinical trial","volume":"42","author":"Zaman","year":"2024","journal-title":"Vaccine"},{"key":"ref51","doi-asserted-by":"publisher","first-page":"894206","DOI":"10.3389\/fimmu.2022.894206","article-title":"Oral administration of a Shigella 2aT32-based vaccine expressing Ure B-HspA fusion antigen with and without parenteral rUreB-HspA boost confers protection against Helicobacter pylori in mice model","volume":"13","author":"Zhang","year":"2022","journal-title":"Front. Immunol."},{"key":"ref52","doi-asserted-by":"publisher","first-page":"5871","DOI":"10.1016\/j.vaccine.2016.10.045","article-title":"Real-time stability of a hepatitis E vaccine (Hecolin\u00ae) demonstrated with potency assays and multifaceted physicochemical methods","volume":"34","author":"Zhang","year":"2016","journal-title":"Vaccine"},{"key":"ref53","doi-asserted-by":"publisher","first-page":"19910","DOI":"10.1074\/jbc.M115.649764","article-title":"A comprehensive study of neutralizing antigenic sites on the hepatitis E virus (HEV) capsid by constructing, clustering, and characterizing a tool box","volume":"290","author":"Zhao","year":"2015","journal-title":"J. Biol. Chem."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1512018\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T09:18:12Z","timestamp":1735031892000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1512018\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,24]]},"references-count":53,"alternative-id":["10.3389\/fmicb.2024.1512018"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2024.1512018","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,24]]},"article-number":"1512018"}}