Persistent Immune Dysregulation during Long COVID is Manifested in Antibodies Targeting Envelope and Nucleocapsid Proteins

Persistent Immune Dysregulation during Long COVID is Manifested in Antibodies Targeting Envelope and Nucleocapsid Proteins | 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 Article Persistent Immune Dysregulation during Long COVID is Manifested in Antibodies Targeting Envelope and Nucleocapsid Proteins Jalees Rehman, Marcin Kwissa, Manikannan Mathayan, Satyajeet Salunkhe, and 26 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8302624/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 Long COVID (LC) or Post-Acute Sequelae of SARS-CoV-2 infection (PASC) syndrome represents a widespread health challenge that necessitates the development of novel diagnostic approaches and targeted therapies that can be readily deployed. Immune dysregulation has been reported as one of the hallmarks of LC, but the extent of LC immune dysregulation in patients over time remains unclear. We therefore assessed SARS-CoV-2-specific antibody responses, peripheral immune cell profiles, autoantibody profiles and circulating cytokines for up to 6 months in participants with a SARS-CoV-2 infection who either convalesced or developed LC. Compared to convalescent, LC participants with a broad range of LC phenotypes exhibited persistently elevated IgG titers for SARS-CoV-2 Envelope and Nucleocapsid proteins over the 6 months of study duration. In contrast, the IgG responses to Spike protein were significantly lower in the LC cohort with predominantly IgG1 and IgG3 class-switched bias. Using CyTOF analysis we show elevated numbers of circulating T follicular helper cells (cTFH) and mucosa- associated invariant T cells (MAIT), which also correlated with high anti-Envelope IgG titers. Persistent immune activation was accompanied by augmented serum cytokine profiles with LIF, IL-11, Eotaxin-3, and HMGB-1 in LC participants, who also demonstrated significantly higher rates of autoantibodies. These findings highlight the persistence of immune dysregulation in LC, underscoring the need to explore targeted therapies addressing viral persistence, dysregulated antibody production, and autoimmunity. Biological sciences/Immunology/Adaptive immunity Biological sciences/Microbiology/Virology/SARS-CoV-2 Health sciences/Diseases/Infectious diseases/Viral infection Full Text Additional Declarations Yes there is potential Competing Interest. Jalees Rehman reports receiving grants from the National Institutes of Health during the conduct of the study. Jerry A. Krishnan reports grants from the National Institutes of Health during the conduct of the study; research grants from the American Lung Association, BioVie Pharma, COPD Foundation, and Patient Centered Outcomes Research Institute outside the submitted work; and personal fees from AstraZeneca, Inogen, MedImmune, RespirAI, and Verona Pharma. Grace A McComsey reports grants from the National Institutes of Health during the conduct of this study; research grants from BioVie, Redhill, Genentech, and Pfizer with money paid to institution, outside of the current work; and personal fees as a research consultant for GlaxoSmithKline/ViiV, Gilead, and Merck, and a member of a GlaxoSmithKline DSMB. Michael J. Peluso reports research funding from the National Institutes of Health, PolyBio Research Foundation, and Patient Led Research Collaborative. He has received consulting fees from Gilead Sciences, AstraZeneca, BioVie, Apellis Pharmaceuticals, and BioNTech, travel support from Invivyd, and research support from Aerium Therapeutics and Shionogi, outside the submitted work. All participants consented to participate in the study. 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-8302624","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":569716318,"identity":"f981f169-b17b-4d83-bbc6-af8573de0a39","order_by":0,"name":"Jalees 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02:00:36","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8302624/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8302624/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102295962,"identity":"980f90ae-57d5-4560-8beb-e1e51a0b1f71","added_by":"auto","created_at":"2026-02-10 10:16:26","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6977378,"visible":true,"origin":"","legend":"Article File","description":"","filename":"LongCOVIDmanuscriptfigstablesextdataFigsMK07Dec25.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8302624/v1_covered_8021440a-75fc-446d-b92f-7b2e5b07523f.pdf"}],"financialInterests":"\u003cp\u003e\u003cstrong\u003eYes\u003c/strong\u003e there is potential Competing Interest. Jalees Rehman reports receiving grants from the National Institutes of Health during the conduct of the study. Jerry A. Krishnan reports grants from the National Institutes of Health during the conduct of the study; research grants from the American Lung Association, BioVie Pharma, COPD Foundation, and Patient Centered Outcomes Research Institute outside the submitted work; and personal fees from AstraZeneca, Inogen, MedImmune, RespirAI, and Verona Pharma. Grace A McComsey reports grants from the National Institutes of Health during the conduct of this study; research grants from BioVie, Redhill, Genentech, and Pfizer with money paid to institution, outside of the current work; and personal fees as a research consultant for GlaxoSmithKline/ViiV, Gilead, and Merck, and a member of a GlaxoSmithKline DSMB. Michael J. Peluso reports research funding from the National Institutes of Health, PolyBio Research Foundation, and Patient Led Research Collaborative. He has received consulting fees from Gilead Sciences, AstraZeneca, BioVie, Apellis Pharmaceuticals, and BioNTech, travel support from Invivyd, and research support from Aerium Therapeutics and Shionogi, outside the submitted work.\u003c/p\u003e\n\u003cp\u003eAll participants consented to participate in the study.\u003c/p\u003e","formattedTitle":"Persistent Immune Dysregulation during Long COVID is Manifested in Antibodies\r\nTargeting Envelope and Nucleocapsid Proteins","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"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-8302624/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8302624/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Long COVID (LC) or Post-Acute Sequelae of SARS-CoV-2 infection (PASC) syndrome\r\nrepresents a widespread health challenge that necessitates the development of novel\r\ndiagnostic approaches and targeted therapies that can be readily deployed. Immune\r\ndysregulation has been reported as one of the hallmarks of LC, but the extent of LC\r\nimmune dysregulation in patients over time remains unclear. We therefore assessed\r\nSARS-CoV-2-specific antibody responses, peripheral immune cell profiles, autoantibody\r\nprofiles and circulating cytokines for up to 6 months in participants with a SARS-CoV-2\r\ninfection who either convalesced or developed LC. Compared to convalescent, LC\r\nparticipants with a broad range of LC phenotypes exhibited persistently elevated IgG titers\r\nfor SARS-CoV-2 Envelope and Nucleocapsid proteins over the 6 months of study\r\nduration. In contrast, the IgG responses to Spike protein were significantly lower in the\r\nLC cohort with predominantly IgG1 and IgG3 class-switched bias. Using CyTOF analysis\r\nwe show elevated numbers of circulating T follicular helper cells (cTFH) and mucosa-\r\nassociated invariant T cells (MAIT), which also correlated with high anti-Envelope IgG\r\ntiters. Persistent immune activation was accompanied by augmented serum cytokine\r\nprofiles with LIF, IL-11, Eotaxin-3, and HMGB-1 in LC participants, who also demonstrated\r\nsignificantly higher rates of autoantibodies. These findings highlight the persistence of\r\nimmune dysregulation in LC, underscoring the need to explore targeted therapies\r\naddressing viral persistence, dysregulated antibody production, and autoimmunity.","manuscriptTitle":"Persistent Immune Dysregulation during Long COVID is Manifested in Antibodies\nTargeting Envelope and Nucleocapsid Proteins","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-08 05:26:23","doi":"10.21203/rs.3.rs-8302624/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":"1c51ee46-380c-45d0-9c22-da34689c6ef6","owner":[],"postedDate":"January 8th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":60619753,"name":"Biological sciences/Immunology/Adaptive immunity"},{"id":60619754,"name":"Biological sciences/Microbiology/Virology/SARS-CoV-2"},{"id":60619755,"name":"Health sciences/Diseases/Infectious diseases/Viral infection"}],"tags":[],"updatedAt":"2026-02-06T08:42:10+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-08 05:26:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8302624","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8302624","identity":"rs-8302624","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}