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As an intracellular pathogen, <jats:italic>Salmonella<\/jats:italic> impacts many cellular pathways. However, the intercellular communication mechanism facilitated by host-derived small extracellular vesicles (EVs), such as exosomes, is an overlooked aspect of the host responses to this infection. We used a comprehensive proteome-based network analysis of exosomes derived from <jats:italic>Salmonella<\/jats:italic>-infected macrophages to identify host molecules that are trafficked via these EVs. This analysis predicted that the host-derived small EVs generated during macrophage infection stimulate macrophages and promote activation of T helper 1 (Th1) cells. We identified that exosomes generated during infection contain <jats:italic>Salmonella<\/jats:italic> proteins, including unique antigens previously shown to stimulate protective immune responses against <jats:italic>Salmonella<\/jats:italic> in murine studies. Furthermore, we showed that host EVs formed upon infection stimulate a mucosal immune response against <jats:italic>Salmonella<\/jats:italic> infection when delivered intranasally to BALB\/c mice, a route of antigen administration known to initiate mucosal immunity. Specifically, the administration of these vesicles to animals stimulated the production of anti-<jats:italic>Salmonella<\/jats:italic> IgG antibodies, such as anti-OmpA antibodies. Exosomes also stimulated antigen-specific cell-mediated immunity. In particular, splenic mononuclear cells isolated from mice administered with exosomes derived from <jats:italic>Salmonella<\/jats:italic>-infected antigen-presenting cells increased CD4+ T cells secreting Th1-type cytokines in response to <jats:italic>Salmonella<\/jats:italic> antigens. These results demonstrate that small EVs, formed during infection, contribute to Th1 cell bias in the anti-<jats:italic>Salmonella<\/jats:italic> responses. Collectively, this study helps to unravel the role of host-derived small EVs as vehicles transmitting antigens to induce Th1-type immunity against Gram-negative bacteria. Understanding the EV-mediated defense mechanisms will allow the development of future approaches to combat bacterial infections.<\/jats:p>","DOI":"10.1371\/journal.ppat.1009465","type":"journal-article","created":{"date-parts":[[2021,5,6]],"date-time":"2021-05-06T17:26:21Z","timestamp":1620321981000},"page":"e1009465","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":52,"title":["Antigen-encapsulating host extracellular vesicles derived from Salmonella-infected cells stimulate pathogen-specific Th1-type responses in vivo"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2662-4992","authenticated-orcid":true,"given":"Winnie W.","family":"Hui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0258-5834","authenticated-orcid":true,"given":"Lisa E.","family":"Emerson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beata","family":"Clapp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2206-0664","authenticated-orcid":true,"given":"Austin E.","family":"Sheppe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4755-9253","authenticated-orcid":true,"given":"Jatin","family":"Sharma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"del Castillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4977-4758","authenticated-orcid":true,"given":"Mark","family":"Ou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6933-4138","authenticated-orcid":true,"given":"Gustavo H. B.","family":"Maegawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3355-7174","authenticated-orcid":true,"given":"Carol","family":"Hoffman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2066-6814","authenticated-orcid":true,"given":"Joseph","family":"Larkin, III","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6572-6916","authenticated-orcid":true,"given":"David W.","family":"Pascual","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0459-7716","authenticated-orcid":true,"given":"Mariola 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