JAK/STAT Inhibition Reprograms T Cell Activation and Metabolism in Inflammatory Arthritis Patients

JAK/STAT Inhibition Reprograms T Cell Activation and Metabolism in Inflammatory Arthritis Patients | 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 JAK/STAT Inhibition Reprograms T Cell Activation and Metabolism in Inflammatory Arthritis Patients Viviana Marzaioli, Aenea AI Brugman, Niamh O' Dowd, Achilleas Floudas, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8050605/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract Objectives: Inflammatory arthritis (IA) is a group of autoimmune diseases characterised by joint inflammation and progressive damage, thus impairing the patient’s quality of life. The JAK/STAT pathway inhibitor Tofacitinib has been successfully introduced into the clinic to treat patients with IA, however its direct effect on T cell responses is widely unknown. This study aims to assess the effect of Tofacitinib on T cell activation, polyfunctionality, proliferation and metabolism. Methods: The effect of Tofacitinib on T cells from peripheral blood, synovial fluid and synovial tissue was evaluated with multidimensional flow cytometric analysis. T cell proliferation was assessed by flow cytometry and T cell metabolism was examined by qPCR and Seahorse XF analyser. To investigate the effect of Tofacitinib on T cell polarisation, naïve T cells were differentiated into Th1, Th2 and Th17 with specific cytokine cocktails. Soluble mediators were evaluated by MSD multiplex analysis. Results: Tofacitinib significantly inhibited T helper cell activation as evidenced by a marked reduction in the frequency of PD-1/CD69/CD25 positive cells (p<0.01). Reduced activation was consistent with impairment of pathogenic polyfunctionality of peripheral blood and synovial tissue-derived T cells. The impact of Tofacitinib on T cell plasticity was further substantiated by reduced T cell polarisation towards Th1 (p<0.05), Th2 (p<0.05), Th17 (p<0.05) and a reduction in genes associated with T cell functions. The attenuation of pathogenic T cell responses is linked to metabolic adaptation, with Tofacitinib leading to a switch in metabolic capacity, mainly ascribed to the CD4-CD8 + T cell compartment. Conclusions: Tofacitinib strongly alters T cell responses and potentially limits T cell pathogenicity by decreasing their activation, polyfunctionality, differentiation, and metabolic potential in both the circulation and the joints of inflammatory arthritis patients. Inflammatory arthritis JAK/STAT inhibition T cells polyfunctionality metabolism Full Text Additional Declarations No competing interests reported. Supplementary Files MarzaiolietalSupplementaryFigures.pptx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 13 Jan, 2026 Reviews received at journal 13 Jan, 2026 Reviews received at journal 12 Jan, 2026 Reviewers agreed at journal 16 Dec, 2025 Reviews received at journal 16 Dec, 2025 Reviewers agreed at journal 16 Dec, 2025 Reviewers agreed at journal 15 Dec, 2025 Reviewers agreed at journal 11 Dec, 2025 Reviewers agreed at journal 10 Dec, 2025 Reviewers invited by journal 19 Nov, 2025 Editor assigned by journal 07 Nov, 2025 Submission checks completed at journal 07 Nov, 2025 First submitted to journal 06 Nov, 2025 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-8050605","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":548218411,"identity":"9ace108e-aee1-4b15-97ef-bae0dde03bd6","order_by":0,"name":"Viviana 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Patients","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"inflammation-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"inre","sideBox":"Learn more about [Inflammation Research](http://link.springer.com/journal/11)","snPcode":"11","submissionUrl":"https://submission.nature.com/new-submission/11/3","title":"Inflammation Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer 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The JAK/STAT pathway inhibitor Tofacitinib has been successfully introduced into the clinic to treat patients with IA, however its direct effect on T cell responses is widely unknown. This study aims to assess the effect of Tofacitinib on T cell activation, polyfunctionality, proliferation and metabolism.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMethods: The effect of Tofacitinib on T cells from peripheral blood, synovial fluid and synovial tissue was evaluated with multidimensional flow cytometric analysis. T cell proliferation was assessed by flow cytometry and T cell metabolism was examined by qPCR and Seahorse XF analyser. To investigate the effect of Tofacitinib on T cell polarisation, naïve T cells were differentiated into Th1, Th2 and Th17 with specific cytokine cocktails. Soluble mediators were evaluated by MSD multiplex analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults:\u0026nbsp;Tofacitinib significantly inhibited T helper cell activation as evidenced by a marked reduction in the frequency of PD-1/CD69/CD25 positive cells (p\u0026lt;0.01). Reduced activation was consistent with impairment of pathogenic polyfunctionality of peripheral blood and synovial tissue-derived T cells. The impact of Tofacitinib on T cell plasticity was further substantiated by reduced T cell polarisation towards Th1 (p\u0026lt;0.05), Th2 (p\u0026lt;0.05), Th17 (p\u0026lt;0.05) and a reduction in genes associated with T cell functions. The attenuation of pathogenic T cell responses is linked to metabolic adaptation, with Tofacitinib leading to a switch in metabolic capacity, mainly ascribed to the CD4-CD8 + T cell compartment.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConclusions: Tofacitinib strongly alters T cell responses and potentially limits T cell pathogenicity by decreasing their activation, polyfunctionality, differentiation, and metabolic potential in both the circulation and the joints of inflammatory arthritis patients.\u003c/p\u003e","manuscriptTitle":"JAK/STAT Inhibition Reprograms T Cell Activation and Metabolism in Inflammatory Arthritis Patients","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-27 08:04:18","doi":"10.21203/rs.3.rs-8050605/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-13T16:42:35+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-13T07:43:27+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-12T21:45:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"99780956623327893796846323474001948213","date":"2025-12-16T21:50:16+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-16T16:33:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"117661073930526024239992873497818708688","date":"2025-12-16T07:48:43+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"77873092708792738168795439465345235434","date":"2025-12-15T15:39:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"63173001051777037397607208750146908064","date":"2025-12-11T10:55:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"87423011938461478903336988459215952107","date":"2025-12-11T02:29:41+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-19T16:36:58+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-07T21:11:20+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-07T21:09:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Inflammation Research","date":"2025-11-06T18:06:40+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"inflammation-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"inre","sideBox":"Learn more about [Inflammation Research](http://link.springer.com/journal/11)","snPcode":"11","submissionUrl":"https://submission.nature.com/new-submission/11/3","title":"Inflammation Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"07722624-65f9-4d28-a118-c134c584adba","owner":[],"postedDate":"November 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-29T15:09:08+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-27 08:04:18","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8050605","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8050605","identity":"rs-8050605","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}