{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T20:39:10Z","timestamp":1778791150641,"version":"3.51.4"},"posted":{"date-parts":[[2025,8,8]]},"group-title":"Allergy and Immunology","reference-count":65,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,8,8]],"date-time":"2025-08-08T00:00:00Z","timestamp":1754611200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Combining cerebrospinal fluid B cell receptor and T cell receptor repertoire analysis with transcriptional\/ flow cytometry cellular profiles in hundreds of deeply-phenotyped people with Multiple Sclerosis (pwMS) and controls, we identified intrathecal expansion of anti-viral, cytotoxic, granzymes H\/K (GZMH+\/GZMK+) double positive (DP) CD8+ T cells that recognize EBV epitopes in pwMS. DP CD8+ T cells are activated and expanded by, and kill autologous, EBV-infected CSF B cell lines in-vitro. Correlations of surrogate transcriptional profiles with clinical and imaging outcomes infer a beneficial role for EBV-targeting DP CD8+ T cells, as untreated pwMS with proportionally higher DP CD8+ T cells to intrathecal B cells accumulate neurological disability slower. MS therapies also increase ratios of beneficial CD8+ T cell responses to intrathecal B cells, consistent with their ability to inhibit disability progression. This study provides indirect evidence that intrathecal EBV infection participates in disability accumulation in pwMS.<\/jats:p>","DOI":"10.1101\/2025.08.05.25333071","type":"posted-content","created":{"date-parts":[[2025,8,9]],"date-time":"2025-08-09T00:40:32Z","timestamp":1754700032000},"source":"Crossref","is-referenced-by-count":2,"title":["Intrathecally expanded GZMK+\/GZMH+ CD8 T cells targeting EBV antigens may reduce severity of Multiple Sclerosis"],"prefix":"10.64898","author":[{"given":"Shinji","family":"Ashida","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Kosa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francisco","family":"Otaizo-Carrasquero","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dan","family":"Sturdevant","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amirhossein","family":"Shamsaddini","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongmei","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jyoti","family":"Shetty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Craig","family":"Martens","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey I.","family":"Cohen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0959-9430","authenticated-orcid":false,"given":"Bibiana","family":"Bielekova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025081108350693000_2025.08.05.25333071v1.1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0801042105"},{"key":"2025081108350693000_2025.08.05.25333071v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/0198-8859(94)90099-X"},{"key":"2025081108350693000_2025.08.05.25333071v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/80516"},{"key":"2025081108350693000_2025.08.05.25333071v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/346183a0"},{"key":"2025081108350693000_2025.08.05.25333071v1.5","doi-asserted-by":"publisher","DOI":"10.1084\/jem.173.1.19"},{"key":"2025081108350693000_2025.08.05.25333071v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81107-1"},{"key":"2025081108350693000_2025.08.05.25333071v1.7","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.172.6.3893"},{"key":"2025081108350693000_2025.08.05.25333071v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(15)00334-8"},{"key":"2025081108350693000_2025.08.05.25333071v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(15)00313-0"},{"key":"2025081108350693000_2025.08.05.25333071v1.10","doi-asserted-by":"publisher","DOI":"10.1002\/ana.21641"},{"key":"2025081108350693000_2025.08.05.25333071v1.11","doi-asserted-by":"crossref","unstructured":"Owens, G.P. , et al. Pathogenic myelin-specific antibodies in multiple sclerosis target conformational proteolipid protein 1-anchored membrane domains. J Clin Invest 133(2023).","DOI":"10.1172\/JCI162731"},{"key":"2025081108350693000_2025.08.05.25333071v1.12","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1110740"},{"key":"2025081108350693000_2025.08.05.25333071v1.13","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04432-7"},{"key":"2025081108350693000_2025.08.05.25333071v1.14","doi-asserted-by":"publisher","DOI":"10.1002\/ana.22449"},{"key":"2025081108350693000_2025.08.05.25333071v1.15","doi-asserted-by":"crossref","first-page":"992","DOI":"10.1128\/jvi.57.3.992-997.1986","article-title":"Demyelinating lesions due to Theiler\u2019s virus are associated with central nervous system infection","volume":"57","year":"1986","journal-title":"J. Virol"},{"key":"2025081108350693000_2025.08.05.25333071v1.16","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01762-07"},{"key":"2025081108350693000_2025.08.05.25333071v1.17","doi-asserted-by":"publisher","DOI":"10.1126\/science.abj8222"},{"key":"2025081108350693000_2025.08.05.25333071v1.18","doi-asserted-by":"publisher","DOI":"10.1007\/s11481-010-9201-3"},{"key":"2025081108350693000_2025.08.05.25333071v1.19","doi-asserted-by":"publisher","DOI":"10.1177\/1352458511408991"},{"key":"2025081108350693000_2025.08.05.25333071v1.20","doi-asserted-by":"crossref","unstructured":"Serafini, B. , et al. Dysregulated Epstein-Barr virus infection in the multiple sclerosis brain. J Exp Med (2007).","DOI":"10.1084\/jem.20071030"},{"key":"2025081108350693000_2025.08.05.25333071v1.21","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/aws315"},{"key":"2025081108350693000_2025.08.05.25333071v1.22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0192109"},{"key":"2025081108350693000_2025.08.05.25333071v1.23","doi-asserted-by":"crossref","unstructured":"Willis, S.N. , et al. Epstein-Barr virus infection is not a characteristic feature of multiple sclerosis brain. Brain : a journal of neurology (2009).","DOI":"10.1093\/brain\/awp200"},{"key":"2025081108350693000_2025.08.05.25333071v1.24","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2008523117"},{"key":"2025081108350693000_2025.08.05.25333071v1.25","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.2.1296-1307.2005"},{"key":"2025081108350693000_2025.08.05.25333071v1.26","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1093\/femsre\/fuz023","article-title":"Epithelial cell infection by Epstein-Barr virus","volume":"43","year":"2019","journal-title":"FEMS Microbiol Rev"},{"key":"2025081108350693000_2025.08.05.25333071v1.27","doi-asserted-by":"publisher","DOI":"10.1084\/jem.190.4.567"},{"key":"2025081108350693000_2025.08.05.25333071v1.28","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0105434"},{"key":"2025081108350693000_2025.08.05.25333071v1.29","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003220"},{"key":"2025081108350693000_2025.08.05.25333071v1.30","doi-asserted-by":"publisher","DOI":"10.1177\/1352458515588581"},{"key":"2025081108350693000_2025.08.05.25333071v1.31","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201444662"},{"key":"2025081108350693000_2025.08.05.25333071v1.32","doi-asserted-by":"crossref","unstructured":"Serafini, B. , Rosicarelli, B. , Veroni, C. , Mazzola, G.A. & Aloisi, F . Epstein-Barr Virus-Specific CD8 T Cells Selectively Infiltrate the Brain in Multiple Sclerosis and Interact Locally with Virus-Infected Cells: Clue for a Virus-Driven Immunopathological Mechanism. J Virol 93(2019).","DOI":"10.1128\/JVI.00980-19"},{"key":"2025081108350693000_2025.08.05.25333071v1.33","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2017-09-805762"},{"key":"2025081108350693000_2025.08.05.25333071v1.34","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1136\/ard-2023-224421","article-title":"Multimodal repertoire analysis unveils B cell biology in immune-mediated diseases","volume":"82","year":"2023","journal-title":"Ann Rheum Dis"},{"key":"2025081108350693000_2025.08.05.25333071v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/nature22976"},{"key":"2025081108350693000_2025.08.05.25333071v1.36","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2020.12.011"},{"key":"2025081108350693000_2025.08.05.25333071v1.37","doi-asserted-by":"crossref","unstructured":"Schafflick, D. , et al. Integrated single cell analysis of blood and cerebrospinal fluid leukocytes in multiple sclerosis. Nat Commun 11, 247 (2020).","DOI":"10.1038\/s41467-019-14118-w"},{"key":"2025081108350693000_2025.08.05.25333071v1.38","doi-asserted-by":"crossref","unstructured":"Ostkamp, P. , et al. A single-cell analysis framework allows for characterization of CSF leukocytes and their tissue of origin in multiple sclerosis. Sci Transl Med 14, eadc9778 (2022).","DOI":"10.1126\/scitranslmed.adc9778"},{"key":"2025081108350693000_2025.08.05.25333071v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-00948-8"},{"key":"2025081108350693000_2025.08.05.25333071v1.40","doi-asserted-by":"crossref","unstructured":"Muller-Miny, L. , et al. Alemtuzumab treatment exemplifies discordant immune effects of blood and cerebrospinal fluid in multiple sclerosis. J Neuroimmunol 378, 578088 (2023).","DOI":"10.1016\/j.jneuroim.2023.578088"},{"key":"2025081108350693000_2025.08.05.25333071v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/J.CELL.2021.04.048"},{"key":"2025081108350693000_2025.08.05.25333071v1.42","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1401513"},{"key":"2025081108350693000_2025.08.05.25333071v1.43","doi-asserted-by":"crossref","unstructured":"Nakayama, M. , Hori, A. , Toyoura, S. & Yamaguchi, S.I . Shaping of T Cell Functions by Trogocytosis. Cells 10(2021).","DOI":"10.3390\/cells10051155"},{"key":"2025081108350693000_2025.08.05.25333071v1.44","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2010-06-284935"},{"key":"2025081108350693000_2025.08.05.25333071v1.45","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1501504"},{"key":"2025081108350693000_2025.08.05.25333071v1.46","doi-asserted-by":"crossref","unstructured":"Stein, J. , et al. Intrathecal B Cells in MS Have Significantly Greater Lymphangiogenic Potential Compared to B Cells Derived From Non-MS Subjects. Front Neurol 9, 554 (2018).","DOI":"10.3389\/fneur.2018.00554"},{"key":"2025081108350693000_2025.08.05.25333071v1.47","unstructured":"Team, R. RStudio: Integrated Development Environment for R. http:\/\/www.rstudio.com\/ (2020)."},{"key":"2025081108350693000_2025.08.05.25333071v1.48","doi-asserted-by":"publisher","DOI":"10.1212\/01.WNL.0000156155.19270.F8"},{"key":"2025081108350693000_2025.08.05.25333071v1.49","doi-asserted-by":"crossref","unstructured":"Kosa, P. , et al. Development of a Sensitive Outcome for Economical Drug Screening for Progressive Multiple Sclerosis Treatment. Front Neurol 7, 131 (2016).","DOI":"10.3389\/fneur.2016.00131"},{"key":"2025081108350693000_2025.08.05.25333071v1.50","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-023-02187-9"},{"key":"2025081108350693000_2025.08.05.25333071v1.51","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneuroim.2010.08.003"},{"key":"2025081108350693000_2025.08.05.25333071v1.52","doi-asserted-by":"crossref","unstructured":"Mittl, K., et al. Antigen specificity of clonally-enriched CD8+ T cells in multiple sclerosis. bioRxiv (2024).","DOI":"10.1101\/2024.09.07.611010"},{"key":"2025081108350693000_2025.08.05.25333071v1.53","doi-asserted-by":"crossref","first-page":"565957","DOI":"10.3389\/fneur.2020.565957","article-title":"Extensive Healthy Donor Age\/Gender Adjustments and Propensity Score Matching Reveal Physiology of Multiple Sclerosis Through Immunophenotyping","volume":"11","year":"2020","journal-title":"Front Neurol"},{"key":"2025081108350693000_2025.08.05.25333071v1.54","doi-asserted-by":"publisher","DOI":"10.1016\/j.cels.2019.03.003"},{"key":"2025081108350693000_2025.08.05.25333071v1.55","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-29356-8"},{"key":"2025081108350693000_2025.08.05.25333071v1.56","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-0969-x"},{"key":"2025081108350693000_2025.08.05.25333071v1.57","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-021-01142-2"},{"key":"2025081108350693000_2025.08.05.25333071v1.58","doi-asserted-by":"crossref","unstructured":"Miao, Y. , et al. Immune Repertoire Profiling Reveals Its Clinical Application Potential and Triggers for Neuromyelitis Optica Spectrum Disorders. Neurol Neuroimmunol Neuroinflamm 10(2023).","DOI":"10.1212\/NXI.0000000000200134"},{"key":"2025081108350693000_2025.08.05.25333071v1.59","doi-asserted-by":"crossref","unstructured":"Aric A. Hagberg , P.S., Daniel S Chult. Exploring Network Structure, Dynamics, and Function Using NetworkX. Python in Science Conference (2008).","DOI":"10.25080\/TCWV9851"},{"key":"2025081108350693000_2025.08.05.25333071v1.60","doi-asserted-by":"publisher","DOI":"10.1101\/gr.1239303"},{"key":"2025081108350693000_2025.08.05.25333071v1.61","doi-asserted-by":"crossref","unstructured":"Langfelder, P. & Horvath, S . WGCNA: an R package for weighted correlation network analysis. BMC Bioinformatics 9, 559 (2008).","DOI":"10.1186\/1471-2105-9-559"},{"key":"2025081108350693000_2025.08.05.25333071v1.62","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkac947"},{"key":"2025081108350693000_2025.08.05.25333071v1.63","doi-asserted-by":"publisher","DOI":"10.1093\/NAR\/GKAD1025"},{"key":"2025081108350693000_2025.08.05.25333071v1.64","doi-asserted-by":"crossref","unstructured":"Sekhon, J.S . Multivariate and Propensity Score Matching Software with Automated Balance Optimization: The Matching Package for R. Journal of Statistical Software 42(2011).","DOI":"10.18637\/jss.v042.i07"},{"key":"2025081108350693000_2025.08.05.25333071v1.65","doi-asserted-by":"publisher","DOI":"10.1101\/2022.10.14.22281095"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.08.05.25333071","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:42:39Z","timestamp":1768509759000},"score":1,"resource":{"primary":{"URL":"http:\/\/medrxiv.org\/lookup\/doi\/10.1101\/2025.08.05.25333071"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,8]]},"references-count":65,"URL":"https:\/\/doi.org\/10.1101\/2025.08.05.25333071","relation":{},"subject":[],"published":{"date-parts":[[2025,8,8]]},"subtype":"preprint"}}