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While infections with pathogenic viruses are well known to modulate extracellular vesicle (EV) biogenesis and function, the extent to which therapeutic poxviral vectors influence EV secretion by immune cells and thereby affect therapeutic efficacy remains underexplored. In this study, we showed that poxviruses, including the clinically relevant Modified Vaccinia Ankara (MVA), stimulate the secretion of small EVs (sEVs) containing viral proteins and immune\u2010related signatures from peripheral blood mononuclear cells (PBMCs). Using an engineered MVA vector, we demonstrated the transfer of virus\u2010encoded therapeutic payloads to sEVs, including the model ovalbumin (OVA)\u2010derived peptide SIINFEKL presented by the class I major histocompatibility complex (MHC I) and the immune activators interleukin\u201012 (IL\u201012) and CD40 ligand (CD40L). Depending on the isolation method, these sEVs stimulated SIINFEKL\u2010specific CD8\n                    <jats:sup>+<\/jats:sup>\n                    T cells with varying efficiencies in vitro. Remarkably, intravenous injection of these sEVs, but also of the soluble secretome from the same cells, into E.G7\u2010OVA lymphoma\u2013bearing mice reduced tumor growth to an extent comparable to the virus itself. Taken together, our findings indicate that EVs released from immune cells infected with engineered therapeutic poxviruses exert potent antitumor activity. These vesicles represent actionable mediators whose secretion and functionalization can be harnessed to improve viral vector\u2013based immunotherapies, as well as being considered as therapeutic vectors in their own.\n                  <\/jats:p>","DOI":"10.1002\/jev2.70307","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T08:45:26Z","timestamp":1781858726000},"source":"Crossref","is-referenced-by-count":0,"title":["Therapeutic Poxviruses Induce the Secretion of Immunostimulating and Anti\u2010Tumoral Extracellular Vesicles"],"prefix":"10.1002","volume":"15","author":[{"given":"Lucas","family":"Walther","sequence":"first","affiliation":[{"name":"Tumor Biomechanics University of Strasbourg  Strasbourg France"},{"name":"INSERM UMR_S1109 University of Strasbourg  Strasbourg France"},{"name":"Universit\u00e9 de Strasbourg  Strasbourg France"},{"name":"F\u00e9d\u00e9ration de M\u00e9decine Translationnelle de Strasbourg (FMTS) University of Strasbourg  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