{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T16:55:02Z","timestamp":1790614502763,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,24]]},"group-title":"Immunology","reference-count":25,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["Pioneer Fellowship"],"award-info":[{"award-number":["Pioneer Fellowship"]}],"id":[{"id":"https:\/\/ror.org\/00jmfr291","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01AA028761"],"award-info":[{"award-number":["R01AA028761"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Alcohol-associated liver disease (ALD) is a major cause of liver-related morbidity and mortality, yet no targeted, disease-modifying therapy exists and the immune mechanisms driving disease progression remain poorly defined. Acetylcholine-producing cholinergic immune cells have emerged as regulators of homeostasis across tissues and in disease, but their role in alcohol-induced hepatic stress is unknown. Here, we characterized the hepatic cholinergic immune landscape during alcohol-induced stress using flow cytometry and single-cell RNA-sequencing of ChAT-eGFP reporter mice. Alcohol feeding expanded hepatic cholinergic immune cells, with T cells constituting a substantial and dynamic fraction of this population that showed a broad increase in acetylcholine signaling and shifted towards a repressed, anergy-like transcriptional state. CD4 T cells underwent coordinated suppression of cholesterol biosynthesis, while CD8 T cells instead adopted a cytotoxic activation program. Together, these findings identify cholinergic T cell signaling, and cholesterol metabolism within CD4 T cells specifically, as a previously unrecognized adaptive response to alcohol-induced liver stress, nominating candidate therapeutic targets for a disease that is marred by critically limited treatment options.<\/jats:p>","DOI":"10.64898\/2026.09.18.752097","type":"posted-content","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T23:10:12Z","timestamp":1790291412000},"source":"Crossref","is-referenced-by-count":0,"title":["Hepatic cholinergic T cell signaling mediates an adaptive response to alcohol-induced liver stress"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8402-2212","authenticated-orcid":false,"given":"Alexander J.","family":"Knights","sequence":"first","affiliation":[{"name":"Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA"},{"name":"Department of Orthopaedic Surgery, Washington University, St Louis, MO, USA"},{"name":"Center of Regenerative Medicine, Washington University, St Louis, MO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanshan","family":"Liu","sequence":"additional","affiliation":[{"name":"Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evan J.","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, Washington University, St Louis, MO, USA"},{"name":"Center of Regenerative Medicine, Washington University, St Louis, MO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4493-5820","authenticated-orcid":false,"given":"Jun","family":"Wu","sequence":"additional","affiliation":[{"name":"Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA"},{"name":"Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026092808551044000_2026.09.18.752097v1.1","doi-asserted-by":"crossref","unstructured":"Danpanichkul P . The global epidemiology of alcohol-associated liver disease. Hepatol Commun. 2026;10(5).","DOI":"10.1097\/HC9.0000000000000947"},{"issue":"1","key":"2026092808551044000_2026.09.18.752097v1.2","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1111\/apt.16969","article-title":"Current and emerging therapies for acute alcohol-associated hepatitis","volume":"56","year":"2022","journal-title":"Aliment Pharmacol Ther"},{"issue":"3","key":"2026092808551044000_2026.09.18.752097v1.3","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1097\/HEP.0000000000000918","article-title":"Kupffer cell diversity maintains liver function in alcohol-associated liver disease","volume":"81","year":"2025","journal-title":"Hepatology"},{"key":"2026092808551044000_2026.09.18.752097v1.4","doi-asserted-by":"publisher","DOI":"10.1126\/science.aau9072"},{"issue":"14","key":"2026092808551044000_2026.09.18.752097v1.5","doi-asserted-by":"crossref","first-page":"e2212476120","DOI":"10.1073\/pnas.2212476120","article-title":"Cholinergic regulation of vascular endothelial function by human ChAT(+) T cells","volume":"120","year":"2023","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"4","key":"2026092808551044000_2026.09.18.752097v1.6","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1038\/s41590-026-02444-3","article-title":"Lymphocyte-derived cholinergic circuits modulate germinal center output and B cell activation","volume":"27","year":"2026","journal-title":"Nat Immunol"},{"key":"2026092808551044000_2026.09.18.752097v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-018-0032-8"},{"key":"2026092808551044000_2026.09.18.752097v1.8","doi-asserted-by":"crossref","unstructured":"Jun H . Adrenergic-Independent Signaling via CHRNA2 Regulates Beige Fat Activation. Dev Cell. 2020.","DOI":"10.1016\/j.devcel.2020.05.017"},{"key":"2026092808551044000_2026.09.18.752097v1.9","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2020106061"},{"key":"2026092808551044000_2026.09.18.752097v1.10","doi-asserted-by":"crossref","unstructured":"Knights AJ . Transcriptomic plasticity of cholinergic adipose macrophages in the acute thermogenic response. J Biol Chem. 2025;301(12).","DOI":"10.1016\/j.jbc.2025.110925"},{"issue":"9","key":"2026092808551044000_2026.09.18.752097v1.11","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1016\/j.it.2022.07.006","article-title":"Immune cell cholinergic signaling in adipose thermoregulation and immunometabolism","volume":"43","year":"2022","journal-title":"Trends Immunol"},{"issue":"6","key":"2026092808551044000_2026.09.18.752097v1.12","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/j.tcb.2021.11.009","article-title":"CHRNA2: a new paradigm in beige thermoregulation and metabolism","volume":"32","year":"2022","journal-title":"Trends Cell Biol"},{"issue":"1","key":"2026092808551044000_2026.09.18.752097v1.13","doi-asserted-by":"crossref","first-page":"113584","DOI":"10.1016\/j.celrep.2023.113584","article-title":"Subcutaneous white adipose tissue independently regulates burn-induced hypermetabolism via immune-adipose crosstalk","volume":"43","year":"2024","journal-title":"Cell Rep"},{"key":"2026092808551044000_2026.09.18.752097v1.14","doi-asserted-by":"crossref","unstructured":"Zhu K . Chrna2-driven CRE Is Expressed in Beige Adipocytes. Endocrinology. 2024;166(1).","DOI":"10.1210\/endocr\/bqae153"},{"issue":"9","key":"2026092808551044000_2026.09.18.752097v1.15","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.2337\/db25-0781","article-title":"Nicotinic Acetylcholine Receptor Signaling Activates Beige Adipocytes and Mediates Systemic Metabolism","volume":"75","year":"2026","journal-title":"Diabetes"},{"key":"2026092808551044000_2026.09.18.752097v1.16","doi-asserted-by":"crossref","unstructured":"Meng W . The miR-182-5p\/FGF21\/acetylcholine axis mediates the crosstalk between adipocytes and macrophages to promote beige fat thermogenesis. JCI Insight. 2021;6(17).","DOI":"10.1172\/jci.insight.150249"},{"issue":"7","key":"2026092808551044000_2026.09.18.752097v1.17","doi-asserted-by":"crossref","first-page":"e3002728","DOI":"10.1371\/journal.pbio.3002728","article-title":"Signaling through the nicotinic acetylcholine receptor in the liver protects against the development of metabolic dysfunction-associated steatohepatitis","volume":"22","year":"2024","journal-title":"PLoS Biol"},{"key":"2026092808551044000_2026.09.18.752097v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2025.04.002"},{"key":"2026092808551044000_2026.09.18.752097v1.19","doi-asserted-by":"publisher","DOI":"10.1186\/s12859-020-3538-2"},{"key":"2026092808551044000_2026.09.18.752097v1.20","doi-asserted-by":"crossref","first-page":"1664","DOI":"10.3389\/fimmu.2017.01664","article-title":"Cholesterol Metabolism in T Cells","volume":"8","year":"2017","journal-title":"Front Immunol"},{"key":"2026092808551044000_2026.09.18.752097v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2013.032"},{"key":"2026092808551044000_2026.09.18.752097v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/S41587-023-01767-Y"},{"key":"2026092808551044000_2026.09.18.752097v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2013.092"},{"key":"2026092808551044000_2026.09.18.752097v1.24","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506580102"},{"key":"2026092808551044000_2026.09.18.752097v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0619-0"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.18.752097","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:55:24Z","timestamp":1790610924000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.18.752097"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,24]]},"references-count":25,"URL":"https:\/\/doi.org\/10.64898\/2026.09.18.752097","relation":{},"subject":[],"published":{"date-parts":[[2026,9,24]]},"subtype":"preprint"}}