{"paper_id":"373cf80b-adfd-4d04-b9d2-47bf7d456e46","body_text":"Structure-based design of a broadly protective influenza hemagglutinin stem immunogen | 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 Structure-based design of a broadly protective influenza hemagglutinin stem immunogen Lei Deng, De-Jian Liu, Yan-Xia Ru, Shi-Long Zhao, Guo-Huan Yang, and 18 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5117559/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The conserved hemagglutinin (HA) stem region is widely regarded as a highly promising target for the development of a universal influenza vaccine. Through structure-based design and iterative mutagenesis screening, we successfully engineered a soluble trimeric H1 stem-only protein, designated #219, which does not contain heterologous trimerization motifs and thus prevents off-target immune responses. As confirmed by a 2.97 Å-resolution cryo-EM structure, the overall conformation of #219 remains intact. Vaccination of mice with #219 protected against lethal heterologous viral challenges. Specific serum antibodies strongly cross-reacted with HAs of diverse influenza A subtypes, cross-neutralized various H1N1 viruses, and exhibited multiple antibody-effector functions mediating infection-permissive protection. We have developed a promising H1 stem immunogen for broad cross-protection against influenza infection and also proposed an efficacious design strategy for soluble homo-oligomeric proteins. Biological sciences/Immunology/Vaccines/Protein vaccines Biological sciences/Structural biology/Electron microscopy/Cryoelectron microscopy Full Text Additional Declarations Yes there is potential Competing Interest. L.D., D.J.L., and H.P.Z. are inventors on a pending patent application (202310583978.4; PCT/CN2023/078609) that discloses the claimed design strategy and combo mutations for constructing stable hemagglutinin stem trimers. The authors declare no other competing interests. Supplementary Files Supplementalinformation.pdf Supplemental material Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-5117559\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":366928788,\"identity\":\"83e5857f-e165-491f-8caa-12c5f9c8c06d\",\"order_by\":0,\"name\":\"Lei 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