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Ideally, these vaccines also induce virus-specific CD8\u2009+\u2009T cells, which have been identified as an independent correlate of protection. In the present study, we explored the use of an artificial immunogen that comprises of twenty highly conserved influenza virus CD8\u2009+\u2009T cell epitopes with an HLA coverage of 99.5% of the world population. The highly attenuated viral vector Modified Vaccinia virus Ankara (MVA) was used to deliver the artificial poly-epitope sequence (rMVA-PE) and by using T cell lines raised against individual epitopes, we confirmed that the epitopes are liberated from the artificial immunogen. For efficient antigen processing and presentation, the epitopes were separated by spacer sequences. Stimulation of peripheral blood mononuclear cells of HLA-typed blood donors with rMVA-PE resulted in the activation of influenza virus-specific T cell responses. Furthermore, immunization of humanized HLA-A2.1-\/HLA-DR1-transgenic H-2 class I-\/class II-knockout mice (HLA-A*02:01) with rMVA-PE induced influenza virus-specific CD8\u2009+\u2009T cell responses. Thus, rMVA-PE proved to be immunogenic both in vitro and in vivo and constitutes a promising vaccine candidate for the induction of cross-reactive CD8\u2009+\u2009T cell responses that could afford protection against antigenically distinct influenza A viruses (IAV) of various subtypes and species, and is currently considered for further clinical testing.<\/jats:p>","DOI":"10.1038\/s41598-025-95479-9","type":"journal-article","created":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T13:58:43Z","timestamp":1743083923000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Design and evaluation of a poly-epitope based vaccine for the induction of influenza A virus cross-reactive CD8\u2009+\u2009T cell responses"],"prefix":"10.1038","volume":"15","author":[{"given":"Sharmistha","family":"Dam","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alina","family":"Tscherne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leoni","family":"Engels","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerd","family":"Sutter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert D. 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