Muscle macrophage regenerative response after squalene-adjuvanted influenza vaccination drives Th2-skewed response and is reduced with age

Muscle macrophage regenerative response after squalene-adjuvanted influenza vaccination drives Th2-skewed response and is reduced with age | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Muscle macrophage regenerative response after squalene-adjuvanted influenza vaccination drives Th2-skewed response and is reduced with age Moataz Noureddine, Lauren Chang, Farah El-Ayache, Gabriel Laghlali, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5760877/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Squalene-based adjuvants like MF59 and AddaVax induce transient muscle injury, but their downstream mechanisms remain unclear. We show that intramuscular injection of a quadrivalent inactivated influenza vaccine (QIV) with AddaVax triggers a muscle regenerative-like immune response, increasing CX3CR1⁺Ly6C⁺ macrophages in muscle and draining lymph nodes within four days. This promotes a type II-skewed immune response, with elevated IgG1 titers and Th2 cytokines in the lungs after influenza challenge. In aged mice, the macrophage response and type II skewing are diminished. Adoptive transfer of young mice bone marrow-derived macrophages into aged mice at vaccination leads to macrophage antigen uptake, lymph node migration, and type II immune bias. However, this restoration does not improve protection against mismatched viral challenge, suggesting additional age-related immune impairments. These findings highlight the role of macrophage-driven muscle regeneration in adjuvant activity and underscore the need to better understand how muscle damage and repair influence intramuscular vaccine efficacy. Biological sciences/Immunology/Vaccines/Adjuvants Biological sciences/Biotechnology/Applied immunology/Vaccines/Adjuvants Vaccination Muscle regeneration Aging Influenza Macrophages Tregs Full Text Additional Declarations The authors declare potential competing interests as follows: The M.S. laboratory has received unrelated funding support in sponsored research agreements from Phio Pharmaceuticals, 7Hills Pharma, ArgenX NV, Ziphius and Moderna. The other authors declare they have no conflicts of interest. Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5760877","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":407989432,"identity":"e7c813df-5b82-4e82-8d61-4c5aaee5d40f","order_by":0,"name":"Moataz Noureddine","email":"","orcid":"","institution":"Icahn School of Medicine at Mount Sinai","correspondingAuthor":false,"prefix":"","firstName":"Moataz","middleName":"","lastName":"Noureddine","suffix":""},{"id":407989433,"identity":"170f4854-d66f-4c06-9d74-9911dc2bdc56","order_by":1,"name":"Lauren Chang","email":"","orcid":"https://orcid.org/0000-0002-3287-3987","institution":"Icahn School of 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