{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T22:58:45Z","timestamp":1768517925201,"version":"3.49.0"},"posted":{"date-parts":[[2025,3,1]]},"group-title":"Immunology","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,3,1]],"date-time":"2025-03-01T00:00:00Z","timestamp":1740787200000},"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":[[2025,3,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>FOXP3+ natural regulatory T cells (nTregs) promote resolution of inflammation and repair of epithelial damage following viral pneumonia-induced lung injury, thus representing a cellular therapy for patients with acute respiratory distress syndrome (ARDS). Whether in vitro induced Tregs (iTregs), which can be rapidly generated in substantial numbers from conventional T cells, also promote lung recovery is unknown. nTregs require specific DNA methylation patterns maintained by the epigenetic regulator, ubiquitin-like with PHD and RING finger domains 1 (UHRF1). Here, we tested whether iTregs promote recovery following viral pneumonia and whether iTregs require UHRF1 for their pro-recovery function. We found that adoptive transfer of iTregs to mice with influenza virus pneumonia promotes lung recovery and that loss of UHRF1-mediated maintenance DNA methylation in iTregs leads to reduced engraftment and a delayed repair response. Transcriptional and DNA methylation profiling of adoptively transferred UHRF1-deficient iTregs that had trafficked to influenza-injured lungs demonstrated transcriptional instability with gain of effector T cell lineage-defining transcription factors. Strategies to promote the stability of iTregs could be leveraged to further augment their pro-recovery function during viral pneumonia and other causes of ARDS.<\/jats:p>","DOI":"10.1101\/2025.02.25.640199","type":"posted-content","created":{"date-parts":[[2025,3,1]],"date-time":"2025-03-01T14:05:14Z","timestamp":1740837914000},"source":"Crossref","is-referenced-by-count":2,"title":["Maintenance DNA methylation is required for induced regulatory T cell reparative function following viral pneumonia"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1232-3256","authenticated-orcid":false,"given":"Anthony M.","family":"Joudi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0149-3463","authenticated-orcid":false,"given":"Jonathan K.","family":"Gurkan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7629-5648","authenticated-orcid":false,"given":"Qianli","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9939-7401","authenticated-orcid":false,"given":"Manuel A. Torres","family":"Acosta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8857-9943","authenticated-orcid":false,"given":"Kathryn A.","family":"Helmin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7126-0518","authenticated-orcid":false,"given":"Luisa","family":"Morales-Nebreda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7332-6947","authenticated-orcid":false,"given":"Nurbek","family":"Mambetsariev","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9016-3257","authenticated-orcid":false,"given":"Carla Patricia Reyes","family":"Flores","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3384-5299","authenticated-orcid":false,"given":"Hiam","family":"Abdala-Valencia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6947-2271","authenticated-orcid":false,"given":"Elizabeth M.","family":"Steinert","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9532-7051","authenticated-orcid":false,"given":"Samuel E.","family":"Weinberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5775-8427","authenticated-orcid":false,"given":"Benjamin D.","family":"Singer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025030402350543000_2025.02.25.640199v1.1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.155.3.1151"},{"key":"2025030402350543000_2025.02.25.640199v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.05.009"},{"key":"2025030402350543000_2025.02.25.640199v1.3","doi-asserted-by":"publisher","DOI":"10.1172\/JCI36498"},{"key":"2025030402350543000_2025.02.25.640199v1.4","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2012-0198OC"},{"key":"2025030402350543000_2025.02.25.640199v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2014.33"},{"key":"2025030402350543000_2025.02.25.640199v1.6","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2017-0019OC"},{"key":"2025030402350543000_2025.02.25.640199v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.08.021"},{"key":"2025030402350543000_2025.02.25.640199v1.8","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2014-0327OC"},{"key":"2025030402350543000_2025.02.25.640199v1.9","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2019-0399OC"},{"key":"2025030402350543000_2025.02.25.640199v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2024.115056"},{"key":"2025030402350543000_2025.02.25.640199v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-024-01914-w"},{"key":"2025030402350543000_2025.02.25.640199v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.09.010"},{"key":"2025030402350543000_2025.02.25.640199v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.07.030"},{"key":"2025030402350543000_2025.02.25.640199v1.14","doi-asserted-by":"publisher","DOI":"10.1172\/JCI124613"},{"key":"2025030402350543000_2025.02.25.640199v1.15","doi-asserted-by":"crossref","unstructured":"Joudi AM , et al. Epigenetic Control of Regulatory T Cell Stability and Function: Implications for Translation. Frontiers in Immunology. 2022;13.","DOI":"10.3389\/fimmu.2022.861607"},{"key":"2025030402350543000_2025.02.25.640199v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1774"},{"key":"2025030402350543000_2025.02.25.640199v1.17","doi-asserted-by":"publisher","DOI":"10.1002\/eji.200939432"},{"key":"2025030402350543000_2025.02.25.640199v1.18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0148474"},{"key":"2025030402350543000_2025.02.25.640199v1.19","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.182.1.259"},{"key":"2025030402350543000_2025.02.25.640199v1.20","doi-asserted-by":"crossref","unstructured":"Landman S , et al. DNA Methyltransferase Inhibition Promotes Th1 Polarization in Human CD4(+)CD25(high) FOXP3(+) Regulatory T Cells but Does Not Affect Their Suppressive Capacity. J Immunol Res. 2018;2018:4973964.","DOI":"10.1155\/2018\/4973964"},{"key":"2025030402350543000_2025.02.25.640199v1.21","doi-asserted-by":"crossref","unstructured":"Varanasi SK , et al. Azacytidine Treatment Inhibits the Progression of Herpes Stromal Keratitis by Enhancing Regulatory T Cell Function. J Virol. 2017;91(7).","DOI":"10.1128\/JVI.02367-16"},{"key":"2025030402350543000_2025.02.25.640199v1.22","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxx049"},{"key":"2025030402350543000_2025.02.25.640199v1.23","doi-asserted-by":"publisher","DOI":"10.2119\/molmed.2013.00159"},{"key":"2025030402350543000_2025.02.25.640199v1.24","first-page":"488","article-title":"DNA Methyltransferase Inhibitor Promotes Human CD4(+)CD25(h)FOXP3(+) Regulatory T Lymphocyte Induction under Suboptimal TCR Stimulation","volume":"7","year":"2016","journal-title":"Front Immunol"},{"key":"2025030402350543000_2025.02.25.640199v1.25","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2020008477"},{"key":"2025030402350543000_2025.02.25.640199v1.26","doi-asserted-by":"crossref","unstructured":"Gao CA , et al. Machine learning links unresolving secondary pneumonia to mortality in patients with severe pneumonia, including COVID-19. J Clin Invest. 2023;133(12).","DOI":"10.1172\/JCI170682"},{"key":"2025030402350543000_2025.02.25.640199v1.27","doi-asserted-by":"publisher","DOI":"10.1126\/science.1147939"},{"key":"2025030402350543000_2025.02.25.640199v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/nature06397"},{"key":"2025030402350543000_2025.02.25.640199v1.29","doi-asserted-by":"publisher","DOI":"10.1038\/nature12488"},{"key":"2025030402350543000_2025.02.25.640199v1.30","doi-asserted-by":"publisher","DOI":"10.1172\/JCI137712"},{"key":"2025030402350543000_2025.02.25.640199v1.31","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20190550"},{"key":"2025030402350543000_2025.02.25.640199v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/ni1428"},{"key":"2025030402350543000_2025.02.25.640199v1.33","doi-asserted-by":"crossref","unstructured":"Mambetsariev N , et al. FOXP3+ Regulatory T Cells Require TBET to Regulate Activated CD8+ T Cells During Recovery from Influenza Infection. bioRxiv. 2024.","DOI":"10.1101\/2024.05.30.596295"},{"key":"2025030402350543000_2025.02.25.640199v1.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2024.04.002"},{"key":"2025030402350543000_2025.02.25.640199v1.35","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20120822"},{"key":"2025030402350543000_2025.02.25.640199v1.36","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20121849"},{"key":"2025030402350543000_2025.02.25.640199v1.37","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20050855"},{"key":"2025030402350543000_2025.02.25.640199v1.38","doi-asserted-by":"publisher","DOI":"10.1038\/nature10772"},{"key":"2025030402350543000_2025.02.25.640199v1.39","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20091999"},{"key":"2025030402350543000_2025.02.25.640199v1.40","doi-asserted-by":"crossref","unstructured":"Morales-Nebreda L , et al. Aging imparts cell-autonomous dysfunction to regulatory T cells during recovery from influenza pneumonia. JCI Insight. 2021;6(6).","DOI":"10.1172\/jci.insight.141690"},{"key":"2025030402350543000_2025.02.25.640199v1.41","doi-asserted-by":"publisher","DOI":"10.1038\/NATURE13903"},{"key":"2025030402350543000_2025.02.25.640199v1.42","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2015-0387LE"},{"key":"2025030402350543000_2025.02.25.640199v1.43","doi-asserted-by":"publisher","DOI":"10.1038\/nature14112"},{"key":"2025030402350543000_2025.02.25.640199v1.44","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0802535"},{"key":"2025030402350543000_2025.02.25.640199v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.04.007"},{"key":"2025030402350543000_2025.02.25.640199v1.46","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2008.09.018"},{"key":"2025030402350543000_2025.02.25.640199v1.47","doi-asserted-by":"crossref","unstructured":"Jovisic M , et al. Differential roles of regulatory T cells in acute respiratory infections. J Clin Invest. 2023;133(14).","DOI":"10.1172\/JCI170505"},{"key":"2025030402350543000_2025.02.25.640199v1.48","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-31285-5"},{"key":"2025030402350543000_2025.02.25.640199v1.49","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2886"},{"key":"2025030402350543000_2025.02.25.640199v1.50","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2022009619"},{"key":"2025030402350543000_2025.02.25.640199v1.51","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.2009.03901.x"},{"key":"2025030402350543000_2025.02.25.640199v1.52","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2019.12.009"},{"key":"2025030402350543000_2025.02.25.640199v1.53","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0846-z"},{"key":"2025030402350543000_2025.02.25.640199v1.54","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20162152"},{"key":"2025030402350543000_2025.02.25.640199v1.55","doi-asserted-by":"crossref","unstructured":"Torres Acosta MA , et al. AMP-activated protein kinase is necessary for Treg cell functional adaptation to microenvironmental stress. bioRxiv. 2023:2023.2011.2029.568904.","DOI":"10.1101\/2023.11.29.568904"},{"key":"2025030402350543000_2025.02.25.640199v1.56","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506580102"},{"key":"2025030402350543000_2025.02.25.640199v1.57","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2015.12.006"},{"key":"2025030402350543000_2025.02.25.640199v1.58","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.003589"},{"key":"2025030402350543000_2025.02.25.640199v1.59","doi-asserted-by":"crossref","unstructured":"Liu Q , et al. Novel enzyme-based reduced representation method for DNA methylation profiling with low inputs. bioRxiv. 2024:2024.2005.2024.595803.","DOI":"10.1101\/2024.05.24.595803"},{"key":"2025030402350543000_2025.02.25.640199v1.60","doi-asserted-by":"crossref","unstructured":"Walter JM , et al. Multidimensional assessment of alveolar T cells in critically ill patients. JCI Insight. 2018;3(17).","DOI":"10.1172\/jci.insight.123287"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.02.25.640199","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T08:09:41Z","timestamp":1768464581000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.02.25.640199"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,1]]},"references-count":60,"URL":"https:\/\/doi.org\/10.1101\/2025.02.25.640199","relation":{},"subject":[],"published":{"date-parts":[[2025,3,1]]},"subtype":"preprint"}}