{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:14:26Z","timestamp":1785402866276,"version":"3.56.0"},"posted":{"date-parts":[[2025,8,28]]},"group-title":"Immunology","reference-count":61,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,8,28]],"date-time":"2025-08-28T00:00:00Z","timestamp":1756339200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["1R21AI154772"],"award-info":[{"award-number":["1R21AI154772"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5RO1AI029564"],"award-info":[{"award-number":["5RO1AI029564"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5R35CA232109"],"award-info":[{"award-number":["5R35CA232109"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["RO1AI125247"],"award-info":[{"award-number":["RO1AI125247"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Pennsylvania Cure"],"award-info":[{"award-number":["Pennsylvania Cure"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["SAP4100083100"],"award-info":[{"award-number":["SAP4100083100"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  T Regulatory T cells (Tregs) from patients with relapsing-remitting multiple sclerosis (RRMS) exhibit impaired suppressive function, yet the underlying molecular mechanisms remain elusive. Single-cell RNA sequencing (scRNAseq) of\n                  <jats:italic>ex vivo-<\/jats:italic>\n                  sorted Tregs from RRMS patients and matched healthy controls (HCs) revealed down-regulation of type I IFN (IFN) and IL-27 signaling pathways in RRMS Tregs. These Tregs showed reduced expression of IFN-stimulated genes (ISGs) (\n                  <jats:italic>ISG15, MX1, IFITM1, IFI44L, OAS1<\/jats:italic>\n                  ), as well as key mediators of Treg suppressive function (\n                  <jats:italic>LGALS3, CD81, FCRL3, CD7, CSTB<\/jats:italic>\n                  ), all suggesting a key role of decreased IFN signaling in RRMS Treg dysfunction. To therapeutically target IFN signaling pathways and improve Treg suppressive functions, we used cGAMP-loaded microparticles (MPs) to activate the stimulator of IFN genes (STING) in experimental autoimmune encephalomyelitis (EAE). cGAMP-MP treatment ameliorated EAE via induction of Tregs expressing IL-27R, IL-10, TGF-b, and Granzyme B. This effect was abolished in Treg-specific IL-27R (Treg\n                  <jats:sup>\u0394Il27ra<\/jats:sup>\n                  ) knockout mice, confirming that IL-27 signaling is essential for Treg suppression.\n                <\/jats:p>\n                <jats:p>\n                  <jats:italic>In vitro<\/jats:italic>\n                  IL-27 stimulation of RRMS-derived Tregs restored expression of IFN pathway genes (\n                  <jats:italic>IRF1, IFNGR, IFI16<\/jats:italic>\n                  ) and Treg suppressive genes (\n                  <jats:italic>ICOS, IKZF3, IL7R, TIGIT<\/jats:italic>\n                  ). Thus, we propose that IL-27 pre-stimulation may restore their suppressive function and migration (via\n                  <jats:italic>CCR6, CCR7, S100A11<\/jats:italic>\n                  and\n                  <jats:italic>S1PR4<\/jats:italic>\n                  ) to the central nervous system (CNS) in future clinical trials.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>\n                    Several studies have reported a role for type I IFN and IL-27 signaling in the induction of suppressive Tregs in autoimmune diseases. We report that RRMS Tregs have decreased expression of type I IFN and IL-27 signalling-related genes in comparison to HCs. The animal model of MS (EAE) was successfully treated with cGAMP-MPs, which, via induction of type I IFN, IL-27 and IL-10, restored Treg suppressive function. A scRNAseq study of Tregs from MS patients revealed that IL-27\n                    <jats:italic>in vitro<\/jats:italic>\n                    stimulation normalized the expression of type I IFN genes and Treg suppressive genes. We propose that IL-27 pre-treatment may enhance Treg suppressive function and migration to the CNS in future clinical trials.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.08.26.671836","type":"posted-content","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T12:25:14Z","timestamp":1756815914000},"source":"Crossref","is-referenced-by-count":0,"title":["Targeting Impaired Type I Interferon\u2013IL-27 Signaling Rescues T Regulatory Cell Suppressive Function in Relapsing-Remitting Multiple Sclerosis"],"prefix":"10.64898","author":[{"given":"Manal","family":"Elzoheiry","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Seyedsadr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Wrobel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sowmya Shree","family":"Gopal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ezgi","family":"Kasimoglu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulien","family":"Baeten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ibrahim","family":"Hamad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mia Rose","family":"Gleason","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8509-2415","authenticated-orcid":false,"given":"Bieke","family":"Broux","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenny","family":"Ting","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Booki","family":"Min","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silva","family":"Markovic-Plese","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025090205250595000_2025.08.26.671836v1.1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20031579"},{"key":"2025090205250595000_2025.08.26.671836v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-018-0051-0"},{"key":"2025090205250595000_2025.08.26.671836v1.3","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1302580"},{"key":"2025090205250595000_2025.08.26.671836v1.4","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2001317"},{"key":"2025090205250595000_2025.08.26.671836v1.5","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1800898"},{"key":"2025090205250595000_2025.08.26.671836v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1703100114"},{"key":"2025090205250595000_2025.08.26.671836v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2015.45"},{"key":"2025090205250595000_2025.08.26.671836v1.8","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/s40259-020-00462-7","article-title":"Administration of CD4(+)CD25(high)CD127(-)FoxP3(+) Regulatory T Cells for Relapsing-Remitting Multiple Sclerosis: A Phase 1 Study","volume":"35","year":"2021","journal-title":"BioDrugs"},{"key":"2025090205250595000_2025.08.26.671836v1.9","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.adm8859"},{"key":"2025090205250595000_2025.08.26.671836v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(17)30470-2"},{"key":"2025090205250595000_2025.08.26.671836v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-24213-6"},{"key":"2025090205250595000_2025.08.26.671836v1.12","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/j.immuni.2022.01.011","article-title":"Interferon-beta acts directly on T cells to prolong allograft survival by enhancing regulatory T cell induction through Foxp3 acetylation","volume":"55","year":"2022","journal-title":"Immunity"},{"key":"2025090205250595000_2025.08.26.671836v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.cytogfr.2014.10.012"},{"key":"2025090205250595000_2025.08.26.671836v1.14","doi-asserted-by":"crossref","unstructured":"M. Chiarini et al., Simultaneous quantification of natural and inducible regulatory T-cell subsets during interferon-beta therapy of multiple sclerosis patients. J Transl Med 18, 169 (2020).","DOI":"10.1186\/s12967-020-02329-5"},{"key":"2025090205250595000_2025.08.26.671836v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2017.10.003"},{"key":"2025090205250595000_2025.08.26.671836v1.16","doi-asserted-by":"crossref","first-page":"e1221","DOI":"10.1002\/cti2.1221","article-title":"ISG15 protects human Tregs from interferon alpha-induced contraction in a cell-intrinsic fashion","volume":"9","year":"2020","journal-title":"Clin Transl Immunology"},{"key":"2025090205250595000_2025.08.26.671836v1.17","doi-asserted-by":"crossref","unstructured":"P. Baeten , et al., Rapamycin rescues loss of function in blood-brain barrier-interacting Tregs. JCI Insight 9 (2024).","DOI":"10.1172\/jci.insight.167457"},{"key":"2025090205250595000_2025.08.26.671836v1.18","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-15-1021"},{"key":"2025090205250595000_2025.08.26.671836v1.19","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1401803"},{"key":"2025090205250595000_2025.08.26.671836v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.02.012"},{"key":"2025090205250595000_2025.08.26.671836v1.21","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1402381"},{"key":"2025090205250595000_2025.08.26.671836v1.22","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.01883"},{"key":"2025090205250595000_2025.08.26.671836v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07018-y"},{"key":"2025090205250595000_2025.08.26.671836v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2928"},{"key":"2025090205250595000_2025.08.26.671836v1.25","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1038\/cmi.2013.38","article-title":"CD52 as both a marker and an effector molecule of T cells with regulatory action: Identification of novel regulatory T cells","volume":"10","year":"2013","journal-title":"Cell Mol Immunol"},{"key":"2025090205250595000_2025.08.26.671836v1.26","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1301926"},{"key":"2025090205250595000_2025.08.26.671836v1.27","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0902005"},{"key":"2025090205250595000_2025.08.26.671836v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2024.08.008"},{"key":"2025090205250595000_2025.08.26.671836v1.29","doi-asserted-by":"crossref","unstructured":"M. Gomez-Herranz , J. Taylor , R. D. Sloan , IFITM proteins: Understanding their diverse roles in viral infection, cancer, and immunity. J Biol Chem 299, 102741 (2023).","DOI":"10.1016\/j.jbc.2022.102741"},{"key":"2025090205250595000_2025.08.26.671836v1.30","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.24669"},{"key":"2025090205250595000_2025.08.26.671836v1.31","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.adp1720"},{"key":"2025090205250595000_2025.08.26.671836v1.32","doi-asserted-by":"publisher","DOI":"10.1101\/gad.5.5.868"},{"key":"2025090205250595000_2025.08.26.671836v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2006.05.006"},{"key":"2025090205250595000_2025.08.26.671836v1.34","first-page":"3100","article-title":"Treg Heterogeneity, Function, and Homeostasis","volume":"10","year":"2019","journal-title":"Front Immunol"},{"key":"2025090205250595000_2025.08.26.671836v1.35","doi-asserted-by":"crossref","unstructured":"L. Han et al., Interleukin 32 Promotes Foxp3(+) Treg Cell Development and CD8(+) T Cell Function in Human Esophageal Squamous Cell Carcinoma Microenvironment. Front Cell Dev Biol 9, 704853 (2021).","DOI":"10.3389\/fcell.2021.704853"},{"key":"2025090205250595000_2025.08.26.671836v1.36","doi-asserted-by":"crossref","unstructured":"Z. Chen et al., PNOC Expressed by B Cells in Cholangiocarcinoma Was Survival Related and LAIR2 Could Be a T Cell Exhaustion Biomarker in Tumor Microenvironment: Characterization of Immune Microenvironment Combining Single-Cell and Bulk Sequencing Technology. Front Immunol 12, 647209 (2021).","DOI":"10.3389\/fimmu.2021.647209"},{"key":"2025090205250595000_2025.08.26.671836v1.37","doi-asserted-by":"publisher","DOI":"10.1038\/nri2889"},{"key":"2025090205250595000_2025.08.26.671836v1.38","doi-asserted-by":"crossref","first-page":"16330","DOI":"10.3390\/ijms160716330","article-title":"Interferon-Beta Therapy of Multiple Sclerosis Patients Improves the Responsiveness of T Cells for Immune Suppression by Regulatory T Cells","volume":"16","year":"2015","journal-title":"Int J Mol Sci"},{"key":"2025090205250595000_2025.08.26.671836v1.39","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006985"},{"key":"2025090205250595000_2025.08.26.671836v1.40","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1016\/j.jaci.2021.05.045","article-title":"Loss of regulatory capacity in Treg cells following rhinovirus infection","volume":"148","year":"2021","journal-title":"J Allergy Clin Immunol"},{"key":"2025090205250595000_2025.08.26.671836v1.41","doi-asserted-by":"crossref","unstructured":"S. Tanwar et al., Type I IFN signaling in T regulatory cells modulates chemokine production and myeloid derived suppressor cells trafficking during EAE. J Autoimmun 115, 102525 (2020).","DOI":"10.1016\/j.jaut.2020.102525"},{"key":"2025090205250595000_2025.08.26.671836v1.42","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1500036"},{"key":"2025090205250595000_2025.08.26.671836v1.43","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1301093"},{"key":"2025090205250595000_2025.08.26.671836v1.44","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2012.03.042"},{"key":"2025090205250595000_2025.08.26.671836v1.45","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.179.5.3268"},{"key":"2025090205250595000_2025.08.26.671836v1.46","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.5.2752"},{"key":"2025090205250595000_2025.08.26.671836v1.47","doi-asserted-by":"publisher","DOI":"10.1038\/s41556-018-0217-x"},{"key":"2025090205250595000_2025.08.26.671836v1.48","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-032713-120236"},{"key":"2025090205250595000_2025.08.26.671836v1.49","doi-asserted-by":"publisher","DOI":"10.1038\/ni.3466"},{"key":"2025090205250595000_2025.08.26.671836v1.50","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1720113115"},{"key":"2025090205250595000_2025.08.26.671836v1.51","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1016\/j.immuni.2017.10.017","article-title":"Regulatory T Cell Migration Is Dependent on Glucokinase-Mediated Glycolysis","volume":"47","year":"2017","journal-title":"Immunity"},{"key":"2025090205250595000_2025.08.26.671836v1.52","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.02716"},{"key":"2025090205250595000_2025.08.26.671836v1.53","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20110278"},{"key":"2025090205250595000_2025.08.26.671836v1.54","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-022-01152-y"},{"key":"2025090205250595000_2025.08.26.671836v1.55","doi-asserted-by":"crossref","unstructured":"H. Zhang et al., An IL-27-Driven Transcriptional Network Identifies Regulators of IL-10 Expression across T Helper Cell Subsets. Cell Rep 33, 108433 (2020).","DOI":"10.1016\/j.celrep.2020.108433"},{"key":"2025090205250595000_2025.08.26.671836v1.56","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0206-z"},{"key":"2025090205250595000_2025.08.26.671836v1.57","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1820039116"},{"key":"2025090205250595000_2025.08.26.671836v1.58","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-019-0232-6"},{"key":"2025090205250595000_2025.08.26.671836v1.59","doi-asserted-by":"publisher","DOI":"10.1038\/nm1564"},{"key":"2025090205250595000_2025.08.26.671836v1.60","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2221007120"},{"key":"2025090205250595000_2025.08.26.671836v1.61","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1900311"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.08.26.671836","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:26:25Z","timestamp":1768469185000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.08.26.671836"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,28]]},"references-count":61,"URL":"https:\/\/doi.org\/10.1101\/2025.08.26.671836","relation":{},"subject":[],"published":{"date-parts":[[2025,8,28]]},"subtype":"preprint"}}