{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T01:42:26Z","timestamp":1791250946806,"version":"4.1.0"},"reference-count":91,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2025,11,18]],"date-time":"2025-11-18T00:00:00Z","timestamp":1763424000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008783","name":"National Research Council of Science & Technology","doi-asserted-by":"crossref","award":["CRC21022"],"award-info":[{"award-number":["CRC21022"]}],"id":[{"id":"10.13039\/501100008783","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100014188","name":"Ministry of Science and ICT, South Korea","doi-asserted-by":"publisher","award":["2710008770; KK-2513-01"],"award-info":[{"award-number":["2710008770; KK-2513-01"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Cryptosporidium parvum<\/jats:italic>\n                      is a waterborne protozoan parasite responsible for diarrheal illness in humans and animals. The lack of effective vaccines and the emergence of antimicrobial resistance underscore the urgent need for novel prophylactic strategies.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A structure-based immunoinformatics approach was used to design a multi-epitope subunit vaccine (MESV) targeting immunogenic regions of\n                      <jats:italic>C. parvum<\/jats:italic>\n                      . Three proteins, Cp15, Cp23, and CpP2, were selected based on antigenicity, non-allergenicity, non-homology with host proteins, and absence of transmembrane domains. B-cell, CD4\n                      <jats:sup>+<\/jats:sup>\n                      , CD8\n                      <jats:sup>+<\/jats:sup>\n                      , and IFN-\u03b3-inducing epitopes were identified and screened for high antigenicity, non-allergenicity, and non-toxicity. To enhance immune recognition, the lipoprotein LprA, a TLR2 agonist, was fused at the N-terminus using an EAAAK linker, and a PADRE sequence was added to improve helper T-cell responses. Linkers were applied to ensure proper epitope separation and processing. Population coverage was analyzed to evaluate the distribution of HLA-restricted epitopes across global populations. Structural modeling and flexibility analysis (CABS-flex) were performed to assess construct stability. Interactions with TLR2 and TLR4 were examined\n                      <jats:italic>via<\/jats:italic>\n                      molecular docking and 100-ns molecular dynamics (MD) simulations, with MM-GBSA used to estimate binding free energies. Immune simulations predicted host immune responses, while codon optimization,\n                      <jats:italic>in silico<\/jats:italic>\n                      cloning, and mRNA secondary structure prediction assessed expression and transcript stability.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The MESV showed strong binding to TLR2 (\u22121328.4 kcal\/mol) and TLR4 (\u22121133.3 kcal\/mol), with MD simulations confirming stable complexes. Immune simulations indicated robust antibody production, T-cell activation, cytokine release, and dendritic cell recruitment. The vaccine demonstrated global HLA population coverage of 95.92%, with favorable expression and mRNA folding profiles.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The MESV construct demonstrated strong immunogenicity, structural stability, and broad population coverage, underscoring its potential as a promising vaccine candidate against\n                      <jats:italic>C. parvum<\/jats:italic>\n                      . Furthermore, experimental validation is warranted to confirm its safety and efficacy.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0334754","type":"journal-article","created":{"date-parts":[[2025,11,18]],"date-time":"2025-11-18T18:27:41Z","timestamp":1763490461000},"page":"e0334754","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["In silico design of a multi-epitope vaccine against Cryptosporidium parvum using structural and immunoinformatics approaches"],"prefix":"10.1371","volume":"20","author":[{"given":"Guneswar","family":"Sethi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Avinash 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