Cross-protective immunity of the hemagglutinin stalk domain presented on the surface of Lactococcus lactis against divergent influenza viruses in mice

Cross-protective immunity of the hemagglutinin stalk domain presented on the surface of Lactococcus lactis against divergent influenza viruses in mice | 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 Research Cross-protective immunity of the hemagglutinin stalk domain presented on the surface of Lactococcus lactis against divergent influenza viruses in mice Han Lei, Tong Gao, Qianhong Cen This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-16035/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 Background Most of the current approaches to influenza vaccine design focus on antibodies against influenza hemagglutinin (HA). However, these influenza vaccines typically provide strain-specific protection against mostly homologous subtypes. There is an urgent need to develop a universal vaccine that confers cross-protection against influenza viruses. Of note, the HA stalk domain (HAs) is a new target for such an influenza vaccine. Results Recombinant L.lactis/pNZ8150-pgsA-HAs constructed in which pgsA was used as an anchor protein and investigated the immunogenicity of HAs, in the mouse model by oral administration without the use of a mucosal adjuvant. Mice were orally vaccinated with L.lactis/pNZ8150-pgsA-HAs, and then produced significant humoral and mucosal immune responses. Importantly, L.lactis/pNZ8150-pgsA-HAs provided significant cross-protection against H5N1, H3N2 or H1N1 virus infection. Conclusions Our data support the hypothesis that HAs presented on the surface of L. lactis can provide cross-protective immunity against influenza A viruses. Taken together, these findings suggest that L.lactis/pNZ8150-pgsA-HAs can be considered an alternative approach to developing a novel universal vaccine during an influenza A pandemic. Applied & Industrial Microbiology L.lactis/pNZ8150-pgsA-HAs Cross-protection Influenza A viruses Figures Figure 1 Figure 2 Figure 3 Full Text 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-16035","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":383957,"identity":"58cb2980-cb88-48d5-8a99-9646e767d656","order_by":1,"name":"Han Lei","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxElEQVRIiWNgGAWjYBACA3YGhgMfGBiYwTweorQA1R6cAdTCQ5IWsHLitZgzMx88bPPnMLu9RALjg7dtDPLmhLRYNrMlHM5tO8zMI5HAbDi3jcFwZwMhhx3mMTic23AbpIVNmreNIcHgAEEt/B8OW/wBa2H/TaQWHobDDGwQW5iJ1MJmcLC37T8zz5mHzZJzzkkYbiCo5Xjz4w8//qQls7cnH/zwpsxGnqAtMJDMwMDYAKQliFQPBHbEKx0Fo2AUjIIRBwBzwDuUGtocKwAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-5156-4258","institution":"Collge of Medicne, Southwest Jiaotong University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Han","middleName":"","lastName":"Lei","suffix":""},{"id":383958,"identity":"fb2de41c-95d6-4cb2-bb85-845f756750db","order_by":2,"name":"Tong Gao","email":"","orcid":"","institution":"College of Medicine, Southwest Jiaotong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tong","middleName":"","lastName":"Gao","suffix":""},{"id":383959,"identity":"66bdf221-2eaa-4199-9118-2ba996bce7eb","order_by":3,"name":"Qianhong Cen","email":"","orcid":"","institution":"College of Medicine, Southwest Jiaotong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qianhong","middleName":"","lastName":"Cen","suffix":""}],"badges":[],"createdAt":"2020-02-29 12:56:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-16035/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-16035/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":596494,"identity":"0a65b62a-307c-4c53-a0ce-2ff1b66a3348","added_by":"auto","created_at":"2020-03-04 21:54:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":252417,"visible":true,"origin":"","legend":"Expression of HAs presented on the L. lactis surface. (a) Schematic description of the full length of HA from the H5N1 strain containing a globular head and stalk domain. SP: Signal peptide; FP: Fusion peptide; TM: Transmembrane domain; CT: Cytoplasmic tail. (b) Schematic diagram of pNZ8150-pgsA-Has. A GS linker was inserted between pgsA and HAs to stabilize the HAs protein expression. (c) Western blot analysis. Lane 1: L.lactis/pNZ8150- pgsA-HAs; Lane 2: L.lactis/pNZ8150-pgsA. (d) Immunofluorescence microscopy assay. L.lactis/pNZ8150-pgsA (left) and L.lactis/pNZ8150-pgsA-HAs (right) (magnification: 1, 000×). (e) Flow cytometric analysis (positive rate: 45.9%).","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/1.png"},{"id":596496,"identity":"435711cb-ffbd-430e-b944-d8ef5a4d1bac","added_by":"auto","created_at":"2020-03-04 21:54:10","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1047799,"visible":true,"origin":"","legend":"Antibody responses detected by ELISA and HI assay. Sera, intestine and upper respiratory washes were collected from the vaccinated mice at day 15 and day 34 after the initial immunization. (a) HA-specific IgG antibody responses in the sera. (b) HA-specific IgA antibody responses in the intestine washes. (c) HA-specific IgA antibody responses in the upper respiratory washes. The data are represented as the mean ± SD. Asterisk indicates significant difference compared with PBS and L.lactis/pNZ8150-pgsA controls (p ˂ 0.05).","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/2.png"},{"id":596497,"identity":"c95e8b3e-5132-45a3-8c61-b910b3bfafdd","added_by":"auto","created_at":"2020-03-04 21:54:10","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":255602,"visible":true,"origin":"","legend":"Cross-protective efficacy against divergent influenza A viruses. The results are expressed in terms of percent body weight (a, b and c), lung virus titers (d, e and f) and percent survival (g, h andi). Two weeks after the last immunization, mice were intranasally infected with 20 μl of 104 EID50 of lethal dose of A/Vietnam/1203/2004(H5N1), A/Beijing/47/1992 (H3N2) and A/California/04/2009(H1N1) (C, F and I) (n=5 / group). The data for lung virus titers are represented as the mean ± SD. Asterisk indicates significant difference compared with PBS and L.lactis/pNZ8150-pgsA controls (p ˂ 0.05).","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/3.jpg"},{"id":791320,"identity":"ca7affc2-6960-4d1f-aa8c-49944a4475b2","added_by":"auto","created_at":"2020-03-31 01:05:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":568095,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/manuscript.pdf"},{"id":605043,"identity":"7fff6654-0102-46a0-9849-60e43a7be956","added_by":"auto","created_at":"2020-03-06 12:01:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":357686,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscripttoMCF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/Manuscript to MCF.pdf"},{"id":596498,"identity":"bd7ec9d8-8822-429b-a64f-9ec381759f75","added_by":"auto","created_at":"2020-03-04 21:54:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":370409,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscripttoMCF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/Manuscript.pdf"},{"id":596495,"identity":"4805b6ed-c827-4f64-8ae2-bf3cfb78f229","added_by":"auto","created_at":"2020-03-04 21:54:10","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":357686,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscripttoMCF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1/Manuscript to MCF.pdf"},{"id":13492010,"identity":"16cb22cc-526e-4fa3-8435-6e94add404ba","added_by":"auto","created_at":"2021-09-16 22:30:01","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":507355,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscripttoMCF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-16035/v1_covered.pdf"}],"financialInterests":"","formattedTitle":"Cross-protective immunity of the hemagglutinin stalk domain presented on the surface of Lactococcus lactis against divergent influenza viruses in mice","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-16035/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"L.lactis/pNZ8150-pgsA-HAs, Cross-protection, Influenza A viruses","lastPublishedDoi":"10.21203/rs.3.rs-16035/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-16035/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground Most of the current approaches to influenza vaccine design focus on antibodies against influenza hemagglutinin (HA). However, these influenza vaccines typically provide strain-specific protection against mostly homologous subtypes. There is an urgent need to develop a universal vaccine that confers cross-protection against influenza viruses. Of note, the HA stalk domain (HAs) is a new target for such an influenza vaccine.\u003c/p\u003e\u003cp\u003eResults Recombinant L.lactis/pNZ8150-pgsA-HAs constructed in which pgsA was used as an anchor protein and investigated the immunogenicity of HAs, in the mouse model by oral administration without the use of a mucosal adjuvant. Mice were orally vaccinated with L.lactis/pNZ8150-pgsA-HAs, and then produced significant humoral and mucosal immune responses. Importantly, L.lactis/pNZ8150-pgsA-HAs provided significant cross-protection against H5N1, H3N2 or H1N1 virus infection.\u003c/p\u003e\u003cp\u003eConclusions Our data support the hypothesis that HAs presented on the surface of L. lactis can provide cross-protective immunity against influenza A viruses. Taken together, these findings suggest that L.lactis/pNZ8150-pgsA-HAs can be considered an alternative approach to developing a novel universal vaccine during an influenza A pandemic.\u003c/p\u003e","manuscriptTitle":"Cross-protective immunity of the hemagglutinin stalk domain presented on the surface of Lactococcus lactis against divergent influenza viruses in mice","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-03-04 21:54:09","doi":"10.21203/rs.3.rs-16035/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"78930d73-ec0c-401d-b2b7-d27387947dc6","owner":[],"postedDate":"March 4th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":66279,"name":"Applied \u0026 Industrial Microbiology"}],"tags":[],"updatedAt":"","versionOfRecord":[],"versionCreatedAt":"2020-03-04 21:54:09","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-16035","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-16035","identity":"rs-16035","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}