{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:43:23Z","timestamp":1791852203029,"version":"4.3.4"},"reference-count":52,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T00:00:00Z","timestamp":1766102400000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Eur J Immunol"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Regulatory T cells (Tregs), defined by the lineage\u2010specific transcription factor FoxP3, are crucial for immune regulation and have been studied extensively in mammals. However, avian Tregs remain poorly characterized, leaving gaps in our understanding of their evolutionary conservation and unique features. In this study, we investigated the phenotype of chicken Tregs to define reliable markers for their identification and characterization. We analyzed CD4\n                    <jats:sup>+<\/jats:sup>\n                    splenocytes sorted into CD25\n                    <jats:sup>negative<\/jats:sup>\n                    , CD25\n                    <jats:sup>low<\/jats:sup>\n                    , and CD25\n                    <jats:sup>high<\/jats:sup>\n                    subpopulations using RNA sequencing.\n                    <jats:italic>FOXP3<\/jats:italic>\n                    and other Treg\u2010associated genes were expressed in both CD25\n                    <jats:sup>low<\/jats:sup>\n                    and CD25\n                    <jats:sup>high<\/jats:sup>\n                    populations, showing that CD25 expression alone is insufficient to distinguish chicken Tregs. To refine the marker profile, we evaluated additional markers, including CTLA\u20104 and GITR. Notably, we describe for the first time a chicken\u2010specific CTLA\u20104 antibody, which uniquely stains CTLA\u20104 exclusively in intracellular (ic) compartments, distinguishing it from mammalian counterparts. Single\u2010cell RNA sequencing further confirmed distinct\n                    <jats:italic>FOXP3<\/jats:italic>\n                    <jats:sup>+<\/jats:sup>\n                    clusters enriched for expression of\n                    <jats:italic>CTLA4<\/jats:italic>\n                    and\n                    <jats:italic>TNFRSF18<\/jats:italic>\n                    (encoding GITR). While CTLA\u20104's ic expression limits usability in functional assays, the combination of CD4\n                    <jats:sup>+<\/jats:sup>\n                    \/CD25\n                    <jats:sup>+<\/jats:sup>\n                    \/CTLA\u20104\n                    <jats:sup>+<\/jats:sup>\n                    \/GITR\n                    <jats:sup>+<\/jats:sup>\n                    represents the most accurate characterization of putative chicken Tregs to date. These findings highlight evolutionary conservation and species\u2010specific differences in Treg markers, providing the foundation for future studies on chicken Treg functionality.\n                  <\/jats:p>","DOI":"10.1002\/eji.70106","type":"journal-article","created":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T14:35:59Z","timestamp":1766154959000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Beyond FoxP3\u2014Identification of a Chicken Regulatory T\u00a0Cell Signature"],"prefix":"10.1002","volume":"55","author":[{"given":"Isabell","family":"Naumann","sequence":"first","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward S.","family":"Ricemeyer","sequence":"additional","affiliation":[{"name":"Bond Life Sciences Center University of Missouri  Columbia Missouri USA"},{"name":"Palaeogenomics Group, Institute of Palaeoanatomy, Domestication Research and the History of Veterinary Medicine Ludwig\u2010Maximilians\u2010Universit\u00e4t  Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Elleder","sequence":"additional","affiliation":[{"name":"Laboratory of Viral and Cellular Genetics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiri","family":"Plachy","sequence":"additional","affiliation":[{"name":"Laboratory of Viral and Cellular Genetics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominik","family":"von La Roche","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Vu\u010dini\u0107","sequence":"additional","affiliation":[{"name":"Laboratory of Genomics and Bioinformatics Institute of Molecular Genetics of the Czech Academy of Sciences  Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas W.","family":"G\u00f6bel","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernd","family":"Kaspers","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon P.","family":"Fr\u00fch","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich Germany"},{"name":"Department of Veterinary Medicine Institute of Virology Freie Universit\u00e4t Berlin  Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7957-7827","authenticated-orcid":false,"given":"Sonja","family":"H\u00e4rtle","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, AG Immunology Ludwig\u2010Maximilians\u2010Universit\u00e4t M\u00fcnchen  Munich 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