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In this study, an adenovirus type 5 vector\u2010based influenza vaccine carrying the hemagglutinin (HA) stem of H1, HA stem of H3, and neuraminidase (NA) of N1 from the influenza virus was constructed. Immune responses were evaluated in mice using various vaccination strategies: prime\u2010only (intramuscular [IM] or intranasal [IN]) and prime\u2010boost (IM\u2009+\u2009IN). Compared with the prime\u2010only strategy, the prime\u2010boost strategy significantly enhanced the systemic immune response, inducing higher levels of antigen\u2010specific IgG, mucosal IgA, and T cell immunity in the spleen and lungs. Furthermore, the IN boosting strategy provided complete protection in mice challenged with the H1N1\u2010PR8, rgH3N2\u2010X31, and rgH5N1\u2010Vietnam viruses, significantly reducing viral loads in the lungs and alleviating lung tissue pathologies. In conclusion, this study elucidates potential avenues for the development and application of universal influenza vaccines using customized mucosal boosting strategies.<\/jats:p>","DOI":"10.1002\/jmv.70326","type":"journal-article","created":{"date-parts":[[2025,3,29]],"date-time":"2025-03-29T23:26:18Z","timestamp":1743290778000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Broad Mucosal and Systemic Immunity in Mice Induced by Intranasal Booster With a Novel Recombinant Adenoviral Based Vaccine Protects Against Divergent Influenza A Virus"],"prefix":"10.1002","volume":"97","author":[{"given":"Jia","family":"Li","sequence":"first","affiliation":[{"name":"Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences Wenzhou Medical University  Wenzhou 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