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Identifying immunogenic peptides with high affinity to different HLA molecules is a promising vaccine strategy. This study used an in silico approach using artificial neural networks to identify\n                    <jats:italic>T. gondii<\/jats:italic>\n                    peptides restricted to HLA-A*02, HLA-A*24, and HLA-B*35 alleles. Proteomes from seven\n                    <jats:italic>T. gondii<\/jats:italic>\n                    strains and transcriptomic data of overexpressed genes from\n                    <jats:italic>T. gondii-<\/jats:italic>\n                    RH in human PBMC were also used. Parasite protein sequences were analyzed with R\n                    <jats:italic>\u2018Epitope Prediction\u2019<\/jats:italic>\n                    library. Peptide candidates were evaluated in the artificial neural networks based on the probabilities of output neurons (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&gt;\u20090.5). The IFN-\u03b3 responses in PBMC from\n                    <jats:italic>T. gondii<\/jats:italic>\n                    seronegative and seropositive individuals were evaluated by ELISpot. Peptides with higher IFN-\u03b3 induction were evaluated to identify cytotoxic response in CD8\n                    <jats:sup>+<\/jats:sup>\n                    T cells (CD107a). In silico analysis identified 36 peptides from\n                    <jats:italic>T. gondii<\/jats:italic>\n                    proteins with predicted affinity to HLA-A*02, A*24, and B*35 alleles. Experiments with PBMCs revealed that a peptide restricted to HLA-A02 (P1: FLFAWITYV) induced a significant increase in IFN-\u03b3-producing cells (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.004). For HLA-A24, a peptide (P8: VFAFAFAFFLI) also induced a significant IFN-\u03b3 response (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.004), while for the HLA-B*35 allele, the P6 peptide (YPIAPSFAM) induced a response that differed significantly from the control (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.05). These peptides induced also a significant percentage of central memory CD8\u2009+\u2009T cells expressing the degranulation marker CD107a (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05). Finally, we identified three\n                    <jats:italic>T. gondii<\/jats:italic>\n                    peptides that induced IFN-\u03b3 response, and a cytotoxic response measured by CD107a expression on CD45RAneg-CD8 cells. These peptides could be considered part of a multi-epitope vaccine against toxoplasmosis in humans.\n                  <\/jats:p>","DOI":"10.1007\/s00430-024-00815-x","type":"journal-article","created":{"date-parts":[[2024,12,31]],"date-time":"2024-12-31T00:27:03Z","timestamp":1735604823000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["In silico identification and ex vivo evaluation of Toxoplasma gondii peptides restricted to HLA-A*02, HLA-A*24 and HLA-B*35 alleles in human PBMC from a Colombian population"],"prefix":"10.1007","volume":"214","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2418-3596","authenticated-orcid":false,"given":"M\u00f3nica","family":"Vargas-Montes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Camila","family":"Valencia-Jaramillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9189-5684","authenticated-orcid":false,"given":"Juan David","family":"Valencia-Hern\u00e1ndez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6472-3329","authenticated-orcid":false,"given":"Jorge Enrique","family":"G\u00f3mez-Mar\u00edn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0331-5556","authenticated-orcid":false,"given":"Ailan Farid","family":"Arenas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6967-165X","authenticated-orcid":false,"given":"N\u00e9stor","family":"Cardona","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,12,31]]},"reference":[{"issue":"1","key":"815_CR1","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1186\/s13071-021-04769-4","volume":"19","author":"JP Dubey","year":"2021","unstructured":"Dubey JP (2021) Outbreaks of clinical toxoplasmosis in humans: five decades of personal experience, perspectives and lessons learned. 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