Highly pathogenic avian influenza virus H5N1 infection in dairy cows confers protective immunity against reinfection

Highly pathogenic avian influenza virus H5N1 infection in dairy cows confers protective immunity against reinfection | 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 Biological Sciences - Article Highly pathogenic avian influenza virus H5N1 infection in dairy cows confers protective immunity against reinfection Yan Zhou, Antonio Facciuolo, Lauren Aubrey, Ulises Barron-Castillo, and 12 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5613077/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 Apr, 2025 Read the published version in Nature Microbiology → Version 1 posted You are reading this latest preprint version Abstract An unprecedented spillover of highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b to dairy cows in March 2024 has affected over 700 US dairy herds in addition to spillback to commercial poultry and transmission to agricultural workers. HPAI H5N1 incursion into cows causes severe mastitis and significant milk production losses. To better understand disease pathogenesis and natural immunity in lactating cows we inoculated the hindquarters of the udder with HPAI H5N1 genotype B3.13 via the teat canal to mimic intra-mammary route of infection. Inoculated cows displayed clinical responses consistent with that observed in affected US dairy herds including generalized decrease in milk yield and localized viral shedding and mastitis in the hindquarters. Following resolution of infection at 31 days post-inoculation, H5N1 virus was inoculated into the unaffected forequarters of the same cows. The secondary inoculation did not result in the clinical manifestations seen upon initial viral challenge. This demonstrated that primary infection of the hindquarters induced natural immunity that conferred complete protection from both mastitis and virus replication and shedding in the forequarters of the udder. This study provides the first experimental report on reinfection demonstrating intra-mammary inoculation can generate H5N1 immunity to completely protect the cow mammary gland. Biological sciences/Microbiology/Virology/Influenza virus Biological sciences/Immunology/Infectious diseases/Influenza virus Highly pathogenic avian influenza H5N1 clade 2.3.4.4b genotype B3.13 dairy cow mastitis immunity Full Text Additional Declarations Yes there is potential Competing Interest. Yanyun Huang, a contributing author, is the CEO of Prairie Diagnostic Services (PDS) and is a board-certified pathologist that provided expert interpretation of the histological slides. PDS provided fee-for-service work that included H&E and IHC staining. Supplementary Files 3ManuscriptV4bFiguresTablesSuppMaterialonlysubmission.pdf Cite Share Download PDF Status: Published Journal Publication published 17 Apr, 2025 Read the published version in Nature Microbiology → 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-5613077","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":389145728,"identity":"5ee3b191-f62c-412f-a766-ba9a1d9dd658","order_by":0,"name":"Yan Zhou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyUlEQVRIie3RsQ6CMBCA4WMnskKI1kdoY8IiDwMLLMRnqAuOrjyGhsQ4lpjoUsNaR8LKoBtsVjenk83E/lOH+3KXFMBk+tUsDuC8X2IM8fhoQsW3hF6uwhqOYVyqtHn0EojDMSJXEXgyiQ8qW/i2AlZgm6jIKLD89CLgwx0i9DhadxRiTcoibYdeE4ISpbdUmuzcKHD1YRHFiKc6Wq3zZFHILlja0mV7jEzqjDVDHk63m7S99eeQzDAyF5+f4SLzOsLxGZPJZPr3nqkUSSu3NX6PAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-4706-1543","institution":"Vaccine and Infectious Disease Organization","correspondingAuthor":true,"prefix":"","firstName":"Yan","middleName":"","lastName":"Zhou","suffix":""},{"id":389145729,"identity":"a09e8367-2d0e-4c4d-b3cf-b814c1093166","order_by":1,"name":"Antonio Facciuolo","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Antonio","middleName":"","lastName":"Facciuolo","suffix":""},{"id":389145730,"identity":"a08a9c60-ea5e-4a21-a3e2-cd712b3a3225","order_by":2,"name":"Lauren Aubrey","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Lauren","middleName":"","lastName":"Aubrey","suffix":""},{"id":389145731,"identity":"a19d6374-c5a8-4455-ae11-7e5543e2c6ed","order_by":3,"name":"Ulises Barron-Castillo","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Ulises","middleName":"","lastName":"Barron-Castillo","suffix":""},{"id":389145732,"identity":"92a4e0f7-b8ae-425b-ac08-ef3f0e893382","order_by":4,"name":"Nathalie Berube","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Nathalie","middleName":"","lastName":"Berube","suffix":""},{"id":389145733,"identity":"8c895532-a574-4ac2-b99a-b1838d5e3711","order_by":5,"name":"Carla Norleen","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Carla","middleName":"","lastName":"Norleen","suffix":""},{"id":389145734,"identity":"ed1b30c6-b006-4a49-800e-824a4e782948","order_by":6,"name":"Shannon McCreary","email":"","orcid":"https://orcid.org/0009-0003-0616-3804","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Shannon","middleName":"","lastName":"McCreary","suffix":""},{"id":389145735,"identity":"1c289542-a3d2-4d90-a5ee-92fcfa1014dc","order_by":7,"name":"Yanyun Huang","email":"","orcid":"","institution":"Prairie Diagnostic Services Inc.","correspondingAuthor":false,"prefix":"","firstName":"Yanyun","middleName":"","lastName":"Huang","suffix":""},{"id":389145736,"identity":"73ce84f3-7b9b-44b5-9599-10a8c1be9834","order_by":8,"name":"Natalia Pessoa","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Natalia","middleName":"","lastName":"Pessoa","suffix":""},{"id":389145737,"identity":"edb5eb2a-57ca-4368-85fd-3b4ae8bcb703","order_by":9,"name":"Leslie Macas Jacome","email":"","orcid":"","institution":"VIDO","correspondingAuthor":false,"prefix":"","firstName":"Leslie","middleName":"Macas","lastName":"Jacome","suffix":""},{"id":389145738,"identity":"f38c7a90-3e07-4c5a-befa-268b48cf19c1","order_by":10,"name":"Samira Mubareka","email":"","orcid":"https://orcid.org/0000-0001-5012-2311","institution":"Sunnybrook Research Institute and University of Toronto","correspondingAuthor":false,"prefix":"","firstName":"Samira","middleName":"","lastName":"Mubareka","suffix":""},{"id":389145739,"identity":"e1dea136-ac4a-4bf3-99d2-593e31a02f57","order_by":11,"name":"Allison McGeer","email":"","orcid":"","institution":"University of Toronto","correspondingAuthor":false,"prefix":"","firstName":"Allison","middleName":"","lastName":"McGeer","suffix":""},{"id":389145740,"identity":"ac1aad07-4299-4b97-b314-10344744b60d","order_by":12,"name":"Yohannes Berhane","email":"","orcid":"","institution":"National Centre for Foreign Animal Disease; 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