Checkpoint inhibition in obesity modulates the intestinal microbiome by disruption of germinal centre formation

Checkpoint inhibition in obesity modulates the intestinal microbiome by disruption of germinal centre formation | 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 Article Checkpoint inhibition in obesity modulates the intestinal microbiome by disruption of germinal centre formation Viviane Schmidt, Roman G. Gerlach, Andrea Deinzer, Jana Petzold, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9576616/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 gut microbiome is an important regulator of the immune system and strongly affected by diet and obesity. Previously, we reported that programmed death-ligand 1 (PD-L1) regulated immune homeostasis in white adipose tissue during diet-induced obesity. Additionally, recent data suggested that checkpoint inhibitiors, such as anti-PD-L1 antibodies, may also alter the host microbiome. Here, we investigated the effect of PD-L1-deficiency or anti-PD-L1 treatment on the intestinal microbiome in lean and obese mice. Within two weeks after starting the high-fat diet, the microbiomes of PD-L1 −/− and wild-type control mice changed and diverged significantly. Mice that were fed with a high-fat diet showed an impaired germinal centre formation and an altered antibody production in the Peyer’s patches. Importantly, similar results were also obtained with wild-type mice that were subjected to immune checkpoint blockade with an anti-PD-L1 antibody during diet-induced obesity. Our results demonstrate that anti-PD-L1 treatment influences the intestinal microbiome during obesity, presumably by impeding the IgA-switch in germinal centres. This finding provides novel insights in our understanding of intestinal inflammation in response to checkpoint inhibitor therapy. PD-L1 checkpoint blockade obesity microbiome germinal centre Figures Figure 3 Full Text Additional Declarations No competing interests reported. Supplementary Files SchmidtsupplementaldataApr30.pdf 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-9576616","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":632720202,"identity":"1a6519ee-747c-468a-add4-3c546e49c345","order_by":0,"name":"Viviane Schmidt","email":"","orcid":"","institution":"Universitätsklinikum Erlangen and Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg","correspondingAuthor":false,"prefix":"","firstName":"Viviane","middleName":"","lastName":"Schmidt","suffix":""},{"id":632720204,"identity":"607c3d46-b61f-445d-b3f6-d30a20331b72","order_by":1,"name":"Roman G. 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Colours show number of statistical tests reporting significantly increased (orange) or decreased (green) abundance at the taxonomic levels indicated. Depicted are the results of three independent experiments. \u003cstrong\u003eb)\u003c/strong\u003e Heatmap of stool samples arranged by: genotype: WT (black) and PD‑L1\u003csup\u003e-/-\u003c/sup\u003e (red) mice; diet: CD (green) and HFD (orange); sex: male (bright blue) and female (pink); and duration of diet: 2 (brown), 4 (blue), 8 (violet) or 14 (orange) weeks. Clusters are highlighted in green (lean cluster L), orange (obese cluster O1, “adaptation”) and blue (obese cluster O2, “obese”). Graph shows results of six to thirteen mice per group (individual columns) from three independent experiments. Composition of the clusters are noted in Supplemental Table 1. \u003cstrong\u003ec)\u003c/strong\u003e Frequency of ILC2s, eosinophils, alternatively activated macrophages and neutrophils in white adipose tissue of male WT (black) or PD‑L1\u003csup\u003e-/-\u003c/sup\u003e (red) mice kept on CD (crosshatched bars) or HFD (filled bars) for 2 weeks. Bars show means+SEM of five to thirteen\u003cstrong\u003e \u003c/strong\u003emice per group\u003cstrong\u003e \u003c/strong\u003efrom five independent experiments. *, p \u0026lt; 0.05; **, p \u0026lt; 0.01; two-way ANOVA or Kruskal-Wallis test. Gating strategies are depicted in Supplemental Fig. 3.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-9576616/v1/a065cd57ecbc3ff0112010d5.png"},{"id":108493806,"identity":"ba819692-b388-4f77-9482-ebfebc963acd","added_by":"auto","created_at":"2026-05-05 10:01:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":787984,"visible":true,"origin":"","legend":"","description":"","filename":"SchmidtsupplementaldataApr30.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9576616/v1/b10d5c8d0259e1ec80276cab.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Checkpoint inhibition in obesity modulates the intestinal microbiome by disruption of germinal centre formation","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":"PD-L1, checkpoint blockade, obesity, microbiome, germinal centre","lastPublishedDoi":"10.21203/rs.3.rs-9576616/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9576616/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe gut microbiome is an important regulator of the immune system and strongly affected by diet and obesity. Previously, we reported that programmed death-ligand 1 (PD-L1) regulated immune homeostasis in white adipose tissue during diet-induced obesity. Additionally, recent data suggested that checkpoint inhibitiors, such as anti-PD-L1 antibodies, may also alter the host microbiome. Here, we investigated the effect of PD-L1-deficiency or anti-PD-L1 treatment on the intestinal microbiome in lean and obese mice.\u003c/p\u003e \u003cp\u003eWithin two weeks after starting the high-fat diet, the microbiomes of PD-L1\u003csup\u003e\u0026minus;/\u0026minus;\u003c/sup\u003e and wild-type control mice changed and diverged significantly. Mice that were fed with a high-fat diet showed an impaired germinal centre formation and an altered antibody production in the Peyer\u0026rsquo;s patches. Importantly, similar results were also obtained with wild-type mice that were subjected to immune checkpoint blockade with an anti-PD-L1 antibody during diet-induced obesity.\u003c/p\u003e \u003cp\u003eOur results demonstrate that anti-PD-L1 treatment influences the intestinal microbiome during obesity, presumably by impeding the IgA-switch in germinal centres. This finding provides novel insights in our understanding of intestinal inflammation in response to checkpoint inhibitor therapy.\u003c/p\u003e","manuscriptTitle":"Checkpoint inhibition in obesity modulates the intestinal microbiome by disruption of germinal centre formation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-05-04 05:57:52","doi":"10.21203/rs.3.rs-9576616/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":"4476916a-5368-4c70-bec9-782bf195b619","owner":[],"postedDate":"May 4th, 2026","published":true,"recentEditorialEvents":[{"type":"checksComplete","content":"","date":"2026-04-30T23:24:22+00:00","index":"","fulltext":""},{"type":"submitted","content":"Microbiome","date":"2026-04-30T11:20:58+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-05-11T17:40:20+00:00","versionOfRecord":[],"versionCreatedAt":"2026-05-04 05:57:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9576616","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9576616","identity":"rs-9576616","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}