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Methods to specifically identify, expand, and manipulate tumor-specific T cells at the rare frequencies found in peripheral blood would enable new immunotherapeutic strategies. Here, we demonstrate an approach to virally transduce polyclonal tumor-reactive T cells across any MHC haplotype and in the absence of knowing the cognate antigen. By generating lentiviral vectors that selectively transduce cells expressing 4-1BB (CD137), a marker of T cell activation, we can transduce antigen-specific T cells with user-defined genetic cargoes that can selectively expand and track individual clonotypes via single-cell sequencing. Anti-4-1BB lentiviruses (4-1BB LVs) encoding therapeutic cargoes can also enhance antigen-specific T cells to extend survival in a xenograft model of human melanoma and transduce tumor-infiltrating T cells from patients with ovarian cancer. Overall, the 4-1BB LV platform targets antigen-specific T cells in a manner agnostic to both the antigen and presenting MHC, with potential applications in adoptive cell therapy manufacturing and TCR identification.<\/jats:p>\n                <jats:sec>\n                  <jats:title>One Sentence Summary<\/jats:title>\n                  <jats:p>Engineered lentiviral vectors targeting 4-1BB selectively activate, expand, and transduce antigen-specific T cells with immunomodulatory cargo.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.11.724165","type":"posted-content","created":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T03:05:19Z","timestamp":1778727919000},"source":"Crossref","is-referenced-by-count":0,"title":["Activation-dependent lentiviruses enable antigen-specific T cell expansion and transduction"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9878-8198","authenticated-orcid":false,"given":"Blake E.","family":"Smith","sequence":"first","affiliation":[{"name":"Koch Institute for Integrative Cancer Research; Cambridge, MA, USA"},{"name":"Department of Immunology, Harvard Medical School; Boston, MA, USA"},{"name":"Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute; Boston, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1107-5640","authenticated-orcid":false,"given":"Lindsey M.","family":"Draper","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of Hematology and Oncology University of California San Francisco; San Francisco, CA, USA"},{"name":"Parker Institute for Cancer Immunotherapy; San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5093-1423","authenticated-orcid":false,"given":"Andrea","family":"Garmilla","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research; Cambridge, MA, USA"},{"name":"Department of Immunology, Harvard Medical School; Boston, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8709-6145","authenticated-orcid":false,"given":"Caleb R.","family":"Perez","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research; Cambridge, MA, USA"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology; Cambridge, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0530-5965","authenticated-orcid":false,"given":"Nishant","family":"Singh","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research; Cambridge, MA, USA"},{"name":"Fletcher Biosciences, Inc.; Cambridge, MA, USA"},{"name":"Ragon Institute of MGH, MIT, and Harvard; Cambridge, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucia T.","family":"Padilla","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research; Cambridge, MA, USA"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology; Cambridge, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4766-9919","authenticated-orcid":false,"given":"Ellen J. 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