In vitro induction of human germinal centre B-cells

In vitro induction of human germinal centre B-cells | 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 In vitro induction of human germinal centre B-cells David Priest, Wataru Ise, James Wing This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6519589/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 During antibody responses, germinal centres (GCs) act as a key site for somatic hypermutation and affinity maturation of B-cells 1 . However, due to their lymphoid tissue localization, experimental manipulation of human germinal centre B-cells has proven difficult, in part due to a lack of an effective method to induce these cells 2 . Here we describe a simple in vitro method, modelling T-cell derived signalling, that allows the conversion of human naive B-cells into in vitro BCL6 + GCB-cells (iGCB). Through side-by-side proteomic and transcriptional single cell analysis of iGCB and GCB from human tonsils, we demonstrate that iGCB-cells have comparable levels of the surface markers, transcription factors, DNA damage and repair molecules, surface glycosylation and trafficking molecules that define human GCB-cells. Using this system, we demonstrate both the signals critical for human GCB formation and a range of interventions that block GCB induction. By tracking plasmablast differentiation, we demonstrate that iGCB derived plasmablasts share markers with tonsil plasmablasts. We anticipate iGCB-cells as a valuable tool for a range of detailed investigations of human B-cell subpopulation formation and behaviour. Biological sciences/Immunology/Adaptive immunity/Humoral immunity/Germinal centres Biological sciences/Immunology/Translational immunology Full Text Additional Declarations The authors declare no competing interests. Supplementary Files ExtendedData.docx 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. We do this by developing innovative software and high quality services for the global research community. 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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-6519589","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":449574024,"identity":"569289f1-3b20-4dab-8403-41fe5b6097fd","order_by":0,"name":"David Priest","email":"","orcid":"","institution":"Human Single Cell Immunology Team, Division of Microbiology and Immunology, Center for Infectious Disease Education and Research (CiDER), Osaka University, Suita, Osaka, Japan","correspondingAuthor":false,"prefix":"","firstName":"David","middleName":"","lastName":"Priest","suffix":""},{"id":449574025,"identity":"3a13e25f-555f-478b-a8b1-eee0d83dbdc8","order_by":1,"name":"Wataru Ise","email":"","orcid":"","institution":"Regulation of Host Defense Team, Division of Microbiology and Immunology, Center for Infectious Disease Education and Research, Osaka University, Osaka, Japan","correspondingAuthor":false,"prefix":"","firstName":"Wataru","middleName":"","lastName":"Ise","suffix":""},{"id":449574023,"identity":"7710fa83-edad-4a55-8fd6-ca3d33de8a2b","order_by":2,"name":"James Wing","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEklEQVRIie2RMUsDMRTHXzg4l1e7RhT7FQyCdKjXr3Il0E107CDHTedSOqdc6WfwI0QCToe3Cl3qNwgIh0gRX+W4xZx1LDS/IST558d7SQA8nj2EKQANMMCjtN7hTZS6hEYZI+r/KvUJA7+VNoJ8KszdZ3mGp+bJwiTpnTzokMN9RJG7DFsUF2Y+WyEejyWHwogcY1KeJUXaraibR9OZrnCIeAUs02wJtxWHUFMU/6W8IGK3IiUZLrtrqvK1Q8EPTQqGpASjnFNjVK5dWZTWdFK5NS55XBg5V+ugP5pJbLuLyDP5jpuIrhK8WTtJrlUZs1dbReei5cXEdptl9arphCYolNOA3s+4cUY7P9Xj8XgOhG/smFmGwRWxFAAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-3462-1003","institution":"Human Single Cell Immunology Team, Division of Microbiology and Immunology, Center for Infectious Disease Education and Research (CiDER), Osaka University, Suita, Osaka, Japan","correspondingAuthor":true,"prefix":"","firstName":"James","middleName":"","lastName":"Wing","suffix":""}],"badges":[],"createdAt":"2025-04-24 10:01:07","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6519589/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-6519589/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":83861725,"identity":"c218c884-9134-4806-8a69-1267812846e4","added_by":"auto","created_at":"2025-06-03 19:40:05","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":6517218,"visible":true,"origin":"","legend":"","description":"","filename":"ExtendedData.docx","url":"https://assets-eu.researchsquare.com/files/rs-6519589/v2/0373f4af9f55dc1e852b27ac.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"In vitro induction of human germinal centre B-cells","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"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":"","lastPublishedDoi":"10.21203/rs.3.rs-6519589/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6519589/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eDuring antibody responses, germinal centres (GCs) act as a key site for somatic hypermutation and affinity maturation of B-cells \u003c/strong\u003e\u003ca href=\"https://paperpile.com/c/kxbrVL/kJPc6\"\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u003c/a\u003e\u003cstrong\u003e. However, due to their lymphoid tissue localization, experimental manipulation of human germinal centre B-cells has proven difficult, in part due to a lack of an effective method to induce these cells \u003c/strong\u003e\u003ca href=\"https://paperpile.com/c/kxbrVL/5wLS1\"\u003e\u003csup\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/sup\u003e\u003c/a\u003e\u003cstrong\u003e. Here we describe a simple \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein vitro\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e method, modelling T-cell derived signalling, that allows the conversion of human naive B-cells into \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein vitro\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e BCL6\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e GCB-cells (iGCB). Through side-by-side proteomic and transcriptional single cell analysis of iGCB and GCB from human tonsils, we demonstrate that iGCB-cells have comparable levels of the surface markers, transcription factors, DNA damage and repair molecules, surface glycosylation and trafficking molecules that define human GCB-cells. Using this system, we demonstrate both the signals critical for human GCB formation and a range of interventions that block GCB induction. By tracking plasmablast differentiation, we demonstrate that iGCB derived plasmablasts share markers with tonsil plasmablasts. We anticipate iGCB-cells as a valuable tool for a range of detailed investigations of human B-cell subpopulation formation and behaviour.\u003c/strong\u003e\u003c/p\u003e","manuscriptTitle":"In vitro induction of human germinal centre B-cells","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2025-06-03 19:40:01","doi":"10.21203/rs.3.rs-6519589/v2","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}},{"code":1,"date":"2025-04-29 09:03:05","doi":"10.21203/rs.3.rs-6519589/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":"b2d56a6d-2542-482c-bb3f-a22bab0a1e86","owner":[],"postedDate":"June 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":47838917,"name":"Biological sciences/Immunology/Adaptive immunity/Humoral immunity/Germinal centres"},{"id":47838918,"name":"Biological sciences/Immunology/Translational immunology"}],"tags":[],"updatedAt":"2025-06-02T15:42:53+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-03 19:40:01","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-6519589","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6519589","identity":"rs-6519589","version":["v2"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}