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This study aims to develop an epitope-based vaccine that targets HIV-1\u2019s key CTL epitopes involved in infection and latency. After studying and carefully checking previous reports and immunoinformatics, a multi-epitope vaccine was designed with strong CTL (Gag p24 and Pol) epitopes. CPP peptide (Tat) and an adjuvant (\u03b2-defensin) were incorporated into the vaccine construct for cell entry and to improve immunogenicity. The designed vaccine was evaluated in terms of antigenicity, allergenicity, safety, various physicochemical properties, solubility, and molecular docking with TLR4 using in silico tools. Immune simulation predicted a significant induction of immune responses. Successful expression and purification of the recombinant protein in\n                    <jats:italic>E. coli<\/jats:italic>\n                    BL21 strain, followed by confirmation by Western blot, demonstrated the feasibility of the vaccine production. Finally, our findings suggest that the designed epitope-based vaccine can induce humoral and cellular immune responses and is a suitable candidate for an HIV-1 therapeutic vaccine to control infection and latency.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-17196-7","type":"journal-article","created":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T13:35:49Z","timestamp":1758807349000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Novel bioinformatics approaches to design epitope-based vaccine against HIV latency by inquiring CTL epitopes and built-in adjuvants"],"prefix":"10.1038","volume":"15","author":[{"given":"Maryam","family":"Mashhadi Abolghasem Shirazi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mina","family":"Hanan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arash","family":"Arashkia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyed Mehdi","family":"Sadat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,9,25]]},"reference":[{"issue":"1","key":"17196_CR1","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1007\/s11904-019-00428-7","volume":"16","author":"RT Veenhuis","year":"2019","unstructured":"Veenhuis, R. 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