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Peptide pulsed Antigen-presenting cells (APCs) are commonly used in cancer immunotherapy to circumvent the defects of tumor cells in processing and presenting antigens to T lymphocytes. However, peptides do not always represent epitopes naturally processed by tumor cells, which might reduce the identification of actual immunogenic antigens. Minigenes encoding concatenated immunogenic tumor epitope sequences offer a promising alternative to select tumor antigens naturally processed and presented to T cells. Hence, using APCs transfected with minigenes might contribute to immunotherapy\u2019s effectiveness, avoiding non-naturally processed epitopes as vaccine candidates. This study evaluates APCs transfected with a minigene construct encoding HLA-A0201-restricted immunogenic antigens to stimulate antigen-specific CD8+ T lymphocytes <jats:italic>in vitro<\/jats:italic>. Artificial APCs (aAPCs) were also designed by co-transfecting the HEK293 cell line with plasmids encoding co-stimulatory molecules (CD80, CD83, CD137L) to assess CD8+ T cell activation efficiency, intracellular cytokine production, cytotoxic activity, activation and exhaustion marker expression. In this study, we successfully implemented a transfection methodology of HEK293 cells with a minigene encoding viral and tumor HLA-A\u2009*\u20090201 epitopes. These cells, used as aAPCs, allow studying the expansion and the phenotype of antigen-specific CD8+ T cells. However, our results indicate that epitope presentation alone is sufficient to activate CD8+ T cells, suggesting that the presence of co-stimulatory molecules may not be necessary for effective T cell activation. Considering that the use of HEK293 cells as aAPCs has yet to be explored and due to their high transfection efficiency with minigenes, the methodology implemented in this work enables their use to identify naturally processed immunogenic neoantigens. We believe our findings can contribute to selecting and designing personalized vaccines based on tumor neoantigens that are useful for cancer immunotherapy.<\/jats:p>","DOI":"10.1371\/journal.pone.0321392","type":"journal-article","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T21:44:33Z","timestamp":1744062273000},"page":"e0321392","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Assessment of antigen immunogenicity formulated in minigenes transfected into antigen-presenting cells"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9709-0426","authenticated-orcid":true,"given":"Mar\u00eda A.","family":"Villota-Alava","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda A.","family":"Alfaro-Marenco","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlos A.","family":"Clavijo-Ram\u00edrez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel A.","family":"Patarroyo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6084-8981","authenticated-orcid":true,"given":"Carlos A.","family":"Parra-L\u00f3pez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,4,7]]},"reference":[{"key":"pone.0321392.ref001","doi-asserted-by":"crossref","first-page":"109821","DOI":"10.1016\/j.biopha.2020.109821","article-title":"Cancer immunotherapy: Pros, cons and beyond.","volume":"124","author":"S Tan","year":"2020","journal-title":"Biomed Pharmacother"},{"issue":"8","key":"pone.0321392.ref002","doi-asserted-by":"crossref","first-page":"1814","DOI":"10.1016\/j.cell.2023.02.039","article-title":"Advances in cancer immunotherapies","volume":"186","author":"OK Dagher","year":"2023","journal-title":"Cell"},{"issue":"8","key":"pone.0321392.ref003","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1038\/s41423-020-0488-6","article-title":"The history and advances in cancer immunotherapy: understanding the characteristics of tumor-infiltrating immune cells and their therapeutic implications","volume":"17","author":"Y Zhang","year":"2020","journal-title":"Cell Mol Immunol"},{"issue":"3","key":"pone.0321392.ref004","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1038\/nrc1815","article-title":"Vaccines for tumour prevention","volume":"6","author":"P-L Lollini","year":"2006","journal-title":"Nat Rev Cancer"},{"issue":"4","key":"pone.0321392.ref005","doi-asserted-by":"crossref","first-page":"706","DOI":"10.3390\/vaccines8040706","article-title":"DC-Based Vaccines for Cancer Immunotherapy","volume":"8","author":"C Fu","year":"2020","journal-title":"Vaccines (Basel)"},{"key":"pone.0321392.ref006","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1007\/978-1-4939-3625-0_15","article-title":"Delivery of Synthetic mRNA Encoding FOXP3 Antigen into Dendritic Cells for Inflammatory Breast Cancer Immunotherapy","author":"GR Devi","year":"2016","journal-title":"Methods Mol Biol"},{"issue":"7662","key":"pone.0321392.ref007","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/nature23003","article-title":"Personalized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer","volume":"547","author":"U Sahin","year":"2017","journal-title":"Nature"},{"issue":"6236","key":"pone.0321392.ref008","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1126\/science.aaa3828","article-title":"Cancer immunotherapy. 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Mortara","year":"2007","journal-title":"Eur J Immunol"},{"issue":"6","key":"pone.0321392.ref026","article-title":"Efficiency of recombinant human TNF in human cancer therapy","volume":"6","author":"F Lejeune","year":"2006","journal-title":"Cancer Immunology"},{"issue":"5","key":"pone.0321392.ref027","doi-asserted-by":"crossref","first-page":"e33","DOI":"10.4110\/in.2018.18.e33","article-title":"Direct antiviral mechanisms of interferon-gamma","volume":"18","author":"S Kang","year":"2018","journal-title":"Immune Network"},{"issue":"4","key":"pone.0321392.ref028","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1038\/7701","article-title":"A human IFNGR1 small deletion hotspot associated with dominant susceptibility to mycobacterial infection","volume":"21","author":"E Jouanguy","year":"1999","journal-title":"Nature Genetics"},{"issue":"2","key":"pone.0321392.ref029","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/S1359-6101(01)00038-7","article-title":"The roles 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