{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T16:49:34Z","timestamp":1786121374440,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,4]]},"group-title":"Microbiology","reference-count":36,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R21AI180874"],"award-info":[{"award-number":["R21AI180874"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21AI176069"],"award-info":[{"award-number":["R21AI176069"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01AI160706"],"award-info":[{"award-number":["R01AI160706"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["75N93021C00014"],"award-info":[{"award-number":["75N93021C00014"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["75N93019C00051"],"award-info":[{"award-number":["75N93019C00051"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32AI007647"],"award-info":[{"award-number":["T32AI007647"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Influenza virus infects the respiratory mucosa, highlighting the importance of mucosal immunity for early protection and transmission control. Here, we evaluated whether intranasal (IN) vaccination with recombinant trimeric hemagglutinin protein from A\/Michigan\/45\/2015 (triHA) formulated with a combined mucosal adjuvant, nanoemulsion plus IVT, an RNA-based RIG-I agonist (NE\/IVT), could protect guinea pigs against heterologous A\/Netherlands\/602\/2009 challenge and reduce viral transmission. To compare mucosal and parenteral immunization, IN triHA\/NE\/IVT was benchmarked against IN triHA alone, IM triHA\/AddaVax (IM triHA\/Advx), and standard IM quadrivalent inactivated influenza vaccine (QIV). We also tested whether IN triHA\/NE\/IVT could boost IM QIV- primed immunity and included animals previously infected with A\/Michigan\/45\/2015 to model pre-existing infection- induced immunity. Transmission was assessed by co-housing na\u00efve sentinels with vaccinated, challenged donors. IN triHA\/NE\/IVT induced systemic humoral responses comparable to IM triHA\/Advx while generating superior nasal mucosal IgA responses. Unexpectedly, IM triHA\/AddaVax also induced detectable, albeit lower, mucosal IgG and IgA, contrasting with prior mouse data and highlighting species-specific differences. IN triHA\/NE\/IVT boosting after IM QIV enhanced serum IgG and mucosal IgA compared with QIV prime-boost alone and increased cross-neutralizing activity against antigenically distinct A\/Victoria\/4897\/2022. Both IN triHA\/NE\/IVT and IN Michigan\/15 prior- infection prevented detectable viral shedding after challenge, and na\u00efve sentinels co-housed with IN triHA\/NE\/IVT- vaccinated donors remained seronegative. Together, these findings support NE\/IVT as a potential mucosal platform capable of inducing robust systemic and mucosal immunity and boosting IM vaccine-primed responses.<\/jats:p>","DOI":"10.64898\/2026.08.03.742639","type":"posted-content","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T04:51:46Z","timestamp":1785905506000},"source":"Crossref","is-referenced-by-count":0,"title":["Adjuvanted mucosal vaccination enhances protection and prevents influenza virus transmission in the guinea pig model"],"prefix":"10.64898","author":[{"given":"Vivian","family":"Yan","sequence":"first","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seok-Chan","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J.","family":"Wiest","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA"},{"name":"Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Laghlali","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Department of Pharmaceutical Sciences, Ghent University, Ghent, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julianne N.","family":"d\u2019Acunzo","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA"},{"name":"Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chinghan","family":"Chung","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA"},{"name":"Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorge","family":"Levican","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farah","family":"El-Ayache","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela T.","family":"Wong","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA"},{"name":"Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan Medical School, Ann Arbor, MI, USA"},{"name":"Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Schotsaert","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA"},{"name":"Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Marc and Jennifer Lipschultz Institute for Precision Immunology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026080621402553000_2026.08.03.742639v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2016.03.085"},{"key":"2026080621402553000_2026.08.03.742639v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2022.11.016"},{"key":"2026080621402553000_2026.08.03.742639v1.3","doi-asserted-by":"crossref","first-page":"24402","DOI":"10.1038\/srep24402","article-title":"Long-Lasting Cross-Protection Against Influenza A by Neuraminidase and M2e-based immunization strategies","volume":"6","year":"2016","journal-title":"Scientific Reports"},{"key":"2026080621402553000_2026.08.03.742639v1.4","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1038\/mi.2012.69","article-title":"Natural and long-lasting cellular immune responses against influenza in the M2e-immune host","volume":"6","year":"2013","journal-title":"Mucosal Immunology"},{"key":"2026080621402553000_2026.08.03.742639v1.5","doi-asserted-by":"crossref","unstructured":"Choi A , Iba\u00f1ez LI , Strohmeier S , Krammer F , Garc\u00eda-Sastre A , Schotsaert M . 2020. Non-sterilizing, Infection-Permissive Vaccination With Inactivated Influenza Virus Vaccine Reshapes Subsequent Virus Infection-Induced Protective Heterosubtypic Immunity From Cellular to Humoral Cross-Reactive Immune Responses. Frontiers in Immunology 11.","DOI":"10.3389\/fimmu.2020.01166"},{"key":"2026080621402553000_2026.08.03.742639v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2022.04.013"},{"key":"2026080621402553000_2026.08.03.742639v1.7","doi-asserted-by":"crossref","first-page":"e2422171122","DOI":"10.1073\/pnas.2422171122","article-title":"Intranasal hemagglutinin protein boosters induce protective mucosal immunity against influenza A viruses in mice","volume":"122","year":"2025","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2026080621402553000_2026.08.03.742639v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2024.10.016"},{"key":"2026080621402553000_2026.08.03.742639v1.9","doi-asserted-by":"crossref","first-page":"2368288","DOI":"10.1080\/21645515.2024.2368288","article-title":"Induction of protective immune responses at respiratory mucosal sites","volume":"20","year":"2024","journal-title":"Hum Vaccin Immunother"},{"key":"2026080621402553000_2026.08.03.742639v1.10","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMp038238"},{"key":"2026080621402553000_2026.08.03.742639v1.11","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1021\/acs.molpharmaceut.0c00315","article-title":"Combined Intranasal Nanoemulsion and RIG-I Activating RNA Adjuvants Enhance Mucosal, Humoral, and Cellular Immunity to Influenza Virus","volume":"18","year":"2021","journal-title":"Molecular Pharmaceutics"},{"key":"2026080621402553000_2026.08.03.742639v1.12","doi-asserted-by":"crossref","unstructured":"Jangra S , Landers JJ , Rathnasinghe R , O\u2019Konek JJ , Janczak KW , Cascalho M , Kennedy AA , Tai AW , Baker JR , Schotsaert M , Wong PT . 2021. A Combination Adjuvant for the Induction of Potent Antiviral Immune Responses for a Recombinant SARS-CoV-2 Protein Vaccine. Frontiers in Immunology 12.","DOI":"10.3389\/fimmu.2021.729189"},{"key":"2026080621402553000_2026.08.03.742639v1.13","doi-asserted-by":"crossref","unstructured":"Jangra S , Landers JJ , Laghlali G , Rathnasinghe R , Warang P , Park S-C , O\u2019Konek JJ , Singh G , Janczak KW , Garc\u00eda-Sastre A , Arya N , Karadag D , Baker JR , Schotsaert M , Wong PT . 2023. Multicomponent intranasal adjuvant for mucosal and durable systemic SARS-CoV-2 immunity in young and aged mice. npj Vaccines 8.","DOI":"10.1038\/s41541-023-00691-1"},{"key":"2026080621402553000_2026.08.03.742639v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/embor.2013.102"},{"key":"2026080621402553000_2026.08.03.742639v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2015.06.002"},{"key":"2026080621402553000_2026.08.03.742639v1.16","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02424-08"},{"key":"2026080621402553000_2026.08.03.742639v1.17","doi-asserted-by":"publisher","DOI":"10.1354\/vp.46-1-138"},{"key":"2026080621402553000_2026.08.03.742639v1.18","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1016\/j.ajpath.2014.11.012","article-title":"Pathogenesis of infection with 2009 pandemic H1N1 influenza virus in isogenic guinea pigs after intranasal or intratracheal inoculation","volume":"185","year":"2015","journal-title":"Am J Pathol"},{"key":"2026080621402553000_2026.08.03.742639v1.19","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1038\/s41541-023-00691-1","article-title":"Multicomponent intranasal adjuvant for mucosal and durable systemic SARS-CoV-2 immunity in young and aged mice","volume":"8","year":"2023","journal-title":"NPJ Vaccines"},{"key":"2026080621402553000_2026.08.03.742639v1.20","doi-asserted-by":"publisher","DOI":"10.1186\/s13567-018-0572-4"},{"key":"2026080621402553000_2026.08.03.742639v1.21","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20020400"},{"key":"2026080621402553000_2026.08.03.742639v1.22","doi-asserted-by":"publisher","DOI":"10.1007\/s10875-012-9768-y"},{"key":"2026080621402553000_2026.08.03.742639v1.23","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2314808120"},{"key":"2026080621402553000_2026.08.03.742639v1.24","doi-asserted-by":"crossref","unstructured":"Kwon D-i , Bhagchandani SH , Ehrenzeller SA , Iwasaki A . 2026. Harnessing mucosal immunity for protective vaccines. Nature Reviews Immunology:1\u201318.","DOI":"10.1038\/s41577-026-01273-7"},{"key":"2026080621402553000_2026.08.03.742639v1.25","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaz5807"},{"key":"2026080621402553000_2026.08.03.742639v1.26","doi-asserted-by":"publisher","DOI":"10.1021\/acs.molpharmaceut.0c00315"},{"key":"2026080621402553000_2026.08.03.742639v1.27","doi-asserted-by":"publisher","DOI":"10.1080\/14760584.2019.1675519"},{"key":"2026080621402553000_2026.08.03.742639v1.28","doi-asserted-by":"crossref","first-page":"2330768","DOI":"10.1080\/21645515.2024.2330768","article-title":"Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses","volume":"20","year":"2024","journal-title":"Hum Vaccin Immunother"},{"key":"2026080621402553000_2026.08.03.742639v1.29","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.12.7481-7486.2001"},{"key":"2026080621402553000_2026.08.03.742639v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-026-01273-7"},{"key":"2026080621402553000_2026.08.03.742639v1.31","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1186\/s12985-015-0448-4","article-title":"Prevalence of gastrointestinal symptoms in patients with influenza, clinical significance, and pathophysiology of human influenza viruses in faecal samples: what do we know?","volume":"12","year":"2015","journal-title":"Virol J"},{"key":"2026080621402553000_2026.08.03.742639v1.32","doi-asserted-by":"publisher","DOI":"10.31486\/toj.20.0086"},{"key":"2026080621402553000_2026.08.03.742639v1.33","first-page":"73","article-title":"Adenovirus: ocular manifestations","volume":"33","year":"2020","journal-title":"Community Eye Health"},{"key":"2026080621402553000_2026.08.03.742639v1.34","doi-asserted-by":"crossref","unstructured":"Zhao C , Pu J . 2022. Influence of Host Sialic Acid Receptors Structure on the Host Specificity of Influenza Viruses. Viruses 14.","DOI":"10.3390\/v14102141"},{"key":"2026080621402553000_2026.08.03.742639v1.35","doi-asserted-by":"crossref","first-page":"e465","DOI":"10.1002\/cpz1.465","article-title":"An In Vitro Microneutralization Assay for Influenza Virus Serology","volume":"2","year":"2022","journal-title":"Curr Protoc"},{"key":"2026080621402553000_2026.08.03.742639v1.36","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1217181"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.08.03.742639","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T04:41:11Z","timestamp":1786077671000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.08.03.742639"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,4]]},"references-count":36,"URL":"https:\/\/doi.org\/10.64898\/2026.08.03.742639","relation":{},"subject":[],"published":{"date-parts":[[2026,8,4]]},"subtype":"preprint"}}