{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T00:23:26Z","timestamp":1783470206826,"version":"3.55.0"},"posted":{"date-parts":[[2025,9,3]]},"group-title":"Immunology","reference-count":76,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T00:00:00Z","timestamp":1756857600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["R01 AI128589"],"award-info":[{"award-number":["R01 AI128589"]}]},{"name":"NIH","award":["R35 CA210043"],"award-info":[{"award-number":["R35 CA210043"]}]},{"name":"NIH","award":["NIAID AI109842"],"award-info":[{"award-number":["NIAID AI109842"]}]},{"name":"NIH","award":["AI040127"],"award-info":[{"award-number":["AI040127"]}]},{"name":"the Tullie and Rickey Families SPARK Awards for Innovations in Immunology at La Jolla Institute"},{"name":"an Astellas Foundation Award for Research on Metabolic Disorders"},{"name":"a Mochida Memorial Foundation Award for Medical and Pharmaceutical Research"},{"DOI":"10.13039\/501100001700","name":"the Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"crossref","award":["JP23K24147"],"award-info":[{"award-number":["JP23K24147"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000875","name":"The Pew Charitable Trusts","doi-asserted-by":"crossref","award":["the Pew Latin-American Fellows Program"],"award-info":[{"award-number":["the Pew Latin-American Fellows Program"]}],"id":[{"id":"10.13039\/100000875","id-type":"DOI","asserted-by":"crossref"}]},{"name":"the California Institute for Regenerative Medicine and a PEW Repatriation Award"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,9,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  T regulatory cells (Treg cells) express the transcription factor FOXP3 and maintain immune homeostasis by attenuating effector responses. Treg cells are prone to lose FOXP3 and convert to pathological \u2018ex-Treg\u2019 cells under conditions of strong or chronic inflammation. One mechanism for loss of FOXP3 expression involves increased DNA methylation of intronic enhancers\n                  <jats:italic>CNS1<\/jats:italic>\n                  and\n                  <jats:italic>CNS2<\/jats:italic>\n                  in the\n                  <jats:italic>Foxp3<\/jats:italic>\n                  locus; these enhancers are maintained in a demethylated state by TET enzymes, 5-methylcytosine (5mC) dioxygenases that generate 5-hydroxymethylcytosine (5hmC) and other oxidized methylcytosines that are essential intermediates in all pathways of DNA demethylation. We previously showed that FOXP3\n                  <jats:sup>+<\/jats:sup>\n                  Treg cells from\n                  <jats:italic>Tet2\/3<\/jats:italic>\n                  -deficient (\n                  <jats:italic>Tet2\/3 DKO<\/jats:italic>\n                  ) mice displayed increased methylation of\n                  <jats:italic>CNS1<\/jats:italic>\n                  and\n                  <jats:italic>CNS2<\/jats:italic>\n                  and converted to FOXP3-negative ex-Treg cells considerably more efficiently than WT Treg cells. Here we extend our previous analysis of\n                  <jats:italic>\n                    Foxp3-Cre Tet2\/3\n                    <jats:sup>fl\/fl<\/jats:sup>\n                  <\/jats:italic>\n                  mice. We classified the mice as DKO- moderate or DKO-severe based on the total number of leukocytes in the spleen and peripheral lymph nodes and investigated the phenotypic and molecular basis for the progressive inflammation occurring in these mice. RNA-seq as well as histological and immunocytochemical analyses showed a striking expansion of T follicular helper (Tfh) cells and plasma cells in\n                  <jats:italic>Tet2\/3 DKO<\/jats:italic>\n                  -severe mice. RNA-seq analyses also revealed increased induction of interferon-stimulated genes (ISGs) in CD4\n                  <jats:sup>+<\/jats:sup>\n                  FOXP3\n                  <jats:sup>-<\/jats:sup>\n                  T cells from these mice, and single-cell (sc) RNA-seq analyses suggested strongly that this was due to skewed differentiation of both\n                  <jats:italic>Tet2\/3 DKO<\/jats:italic>\n                  FOXP3\n                  <jats:sup>+<\/jats:sup>\n                  Treg cells and\n                  <jats:italic>Tet2\/3 DKO<\/jats:italic>\n                  FOXP3\n                  <jats:sup>\u2212<\/jats:sup>\n                  ex-Treg cells into Tfh-like cells. Base-resolution \u201c6-base\u201d sequencing showed the expected loss of 5hmC and increased 5mC in Tfh cells purified from\n                  <jats:italic>Tet2\/3 DKO<\/jats:italic>\n                  -severe mice, and suggested that the observed bias in gene expression patterns could arise both from a direct increase in methylation of essential enhancers stemming from TET deficiency, or because methylation interfered with binding of methylation-sensitive transcriptional regulators including CTCF.\n                <\/jats:p>","DOI":"10.1101\/2025.08.29.673187","type":"posted-content","created":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T13:30:12Z","timestamp":1756906212000},"source":"Crossref","is-referenced-by-count":1,"title":["Loss of TET function in T regulatory cells yields ex-Treg cells biased toward T follicular helper cells, causing autoimmune diseases through autoantibody production"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9342-1300","authenticated-orcid":false,"given":"Kazumasa","family":"Suzuki","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leo J.","family":"Arteaga-Vazquez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno Villalobos","family":"Reveles","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lot","family":"Hern\u00e1ndez-Espinosa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isaac F.","family":"L\u00f3pez-Moyado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atsushi","family":"Onodera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Samaniego-Castruita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ferhat","family":"Ay","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arlet","family":"Lara-Custodio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick 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