{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T01:42:49Z","timestamp":1791250969905,"version":"4.1.0"},"reference-count":92,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T00:00:00Z","timestamp":1738022400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Leptospirosis, caused by <jats:italic>Leptospira interrogans<\/jats:italic>, is a neglected zoonotic disease that poses a significant global health risk to both humans and animals. The rise of antimicrobial resistance and the inefficacy of existing vaccines highlight the urgent need for new preventive strategies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>An immunoinformatics approach was employed to design a multi-epitope subunit vaccine (MESV) against leptospirosis. B-cell, cytotoxic T lymphocyte (CTL), and helper T lymphocyte (HTL) epitopes were selected from five key <jats:italic>Leptospira<\/jats:italic> proteins. These epitopes were fused with a heparin-binding hemagglutinin (HBHA) adjuvant and appropriate linkers to construct the broad-spectrum vaccine. The physicochemical properties of the vaccine were assessed, including antigenicity, immunogenicity, allergenicity, and conservation. The vaccine\u2019s 3D structure was modeled, optimized, and validated. Molecular docking, molecular dynamics simulations, and MM-GBSA analysis were performed to assess the vaccine's binding interactions with Toll-like receptors (TLR2 and TLR4). Immune simulations and <jats:italic>in silico<\/jats:italic> cloning were also conducted to evaluate the vaccine\u2019s immune response and expression potential.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The MESV demonstrated high antigenicity, immunogenicity, non-allergenicity, and conservation across different <jats:italic>Leptospira<\/jats:italic> strains. Population coverage analysis revealed that T-cell epitopes significantly interacted with HLA molecules, covering 95.7% of the global population. Molecular docking showed strong and stable binding with TLR2 and TLR4, with binding energies of -1,357.1 kJ\/mol and -1,163.7 kJ\/mol, respectively. Molecular dynamics simulations and MM-GBSA analysis confirmed the stability of these interactions and accurately calculated the intermolecular binding free energies. Immune simulations indicated robust B and T cell responses, and <jats:italic>in silico<\/jats:italic> cloning demonstrated that the vaccine could be successfully expressed in <jats:italic>E. coli<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>These findings suggest that MESV is a promising candidate for leptospirosis prevention, providing robust immune responses and broad population coverage. However, further <jats:italic>in vivo<\/jats:italic> studies are necessary to validate its efficacy and safety.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2024.1503853","type":"journal-article","created":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T06:45:08Z","timestamp":1738046708000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":15,"title":["Designing a broad-spectrum multi-epitope subunit vaccine against leptospirosis using immunoinformatics and structural approaches"],"prefix":"10.3389","volume":"15","author":[{"given":"Guneswar","family":"Sethi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young Kyu","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su-Cheol","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeong Ho","family":"Hwang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,1,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3083.2010.02505.x","article-title":"Leptospirosis: aspects of innate immunity, immunopathogenesis and immune evasion from the complement system","volume":"73","author":"Fraga","year":"2011","journal-title":"Scand J Immunol"},{"key":"B2","doi-asserted-by":"publisher","first-page":"e0003898","DOI":"10.1371\/journal.pntd.0003898","article-title":"Global morbidity and mortality of leptospirosis: A systematic review","volume":"9","author":"Costa","year":"2015","journal-title":"PloS Negl Trop Dis"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens12091167","article-title":"Leptospirosis and the environment: A review and future directions","volume":"12","author":"Bradley","year":"2023","journal-title":"Pathogens"},{"key":"B4","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1007\/978-3-662-45059-8_5","article-title":"Leptospirosis in humans","volume":"387","author":"Haake","year":"2015","journal-title":"Curr Top Microbiol Immunol"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines12020435","article-title":"Fulminant leptospirosis presenting with rapidly developing acute renal failure and multiorgan failure","volume":"12","author":"Liu","year":"2024","journal-title":"Biomedicines"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1016\/j.prevetmed.2024.106255","article-title":"Estimation of economic losses due to leptospirosis in dairy cattle","volume":"229","author":"Carvalho","year":"2024","journal-title":"Prev Vet Med"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2020.104241","article-title":"Leptospira spp., a genus in the stage of diversity and genomic data expansion","volume":"81","author":"Caimi","year":"2020","journal-title":"Infect Genet Evol"},{"key":"B8","doi-asserted-by":"publisher","first-page":"e0007270","DOI":"10.1371\/journal.pntd.0007270","article-title":"Revisiting the taxonomy and evolution of pathogenicity of the genus Leptospira through the prism of genomics","volume":"13","author":"Vincent","year":"2019","journal-title":"PloS Negl Trop Dis"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens10080943","article-title":"Regional prevalence of intermediate leptospira spp","volume":"10","author":"Abd Rahman","year":"2021","journal-title":"Humans: A Meta-Analysis. 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