{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T02:54:26Z","timestamp":1768272866330,"version":"3.49.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T00:00:00Z","timestamp":1672876800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Autoimmune hepatitis (AIH) is a chronic liver disease caused by a perturbed immune system. The scarcity of short- and long-term immune monitoring of AIH hampered us to comprehend the interaction between immunosuppressive medication and immune homeostasis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and patients<\/jats:title><jats:p>We recruited children with AIH at the time of diagnosis and at the 1st, 3rd, 6th, 12th, 18th, and 24th months of immunosuppression (IS). We also enrolled children with AIH being on IS for &amp;gt;2 years. Children with drug-induced liver injury (DILI), and those receiving tacrolimus after liver transplantation (LT), were enrolled as disease\/IS control subjects. Healthy children (HC) were also recruited. Peripheral blood mononuclear cells (PBMCs) were isolated from all participants. Healthy liver tissue from adult donors and from livers without inflammation were obtained from children with hepatoblastoma. By using flow cytometry, we performed multi-parametric immune profiling of PBMCs and intrahepatic lymphocytes. Additionally, after IS with prednisolone, tacrolimus, rapamycin, or 6-mercaptopurine, we carried out an <jats:italic>in vitro<\/jats:italic> cytokine stimulation assay. Finally, a Lifecodes SSO typing kit was used to type HLA-DRB1 and Luminex was used to analyze the results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Untreated AIH patients had lower total CD8 T-cell frequencies than HC, but these cells were more na\u00efve. While the percentage of na\u00efve regulatory T cells (Tregs) (CD4<jats:sup>+<\/jats:sup>FOXP3<jats:sup>low<\/jats:sup>CD45RA<jats:sup>+<\/jats:sup>) and regulatory B cells (Bregs, CD20<jats:sup>+<\/jats:sup>CD24<jats:sup>+<\/jats:sup>CD38<jats:sup>+<\/jats:sup>) was similar, AIH patients had fewer activated Tregs (CD4<jats:sup>+<\/jats:sup>FOXP3<jats:sup>high<\/jats:sup>CD45RA<jats:bold><jats:sup>-<\/jats:sup><\/jats:bold>) compared to HC. Mucosal-associated-invariant-T-cells (MAIT) were also lower in these patients. Following the initiation of IS, the immune profiles demonstrated fluctuations. Bregs frequency decreased substantially at 1 month and did not recover anymore. Additionally, the frequency of intrahepatic Bregs in treated AIH patients was lower, compared to control livers, DILI, and LT patients. Following <jats:italic>in vitro<\/jats:italic> IS drugs incubation, only the frequency of IL-10-producing total B-cells increased with tacrolimus and 6MP. Lastly, 70% of AIH patients possessed HLA-DR11, whereas HLA-DR03\/DR07\/DR13 was present in only some patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>HLA-DR11 was prominent in our AIH cohort. Activated Tregs and MAIT cell frequencies were lower before IS. Importantly, we discovered a previously unrecognized and long-lasting Bregs scarcity in AIH patients after IS. Tacrolimus and 6MP increased IL-10+ B-cells <jats:italic>in vitro<\/jats:italic>.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2022.1053216","type":"journal-article","created":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T11:46:43Z","timestamp":1672919203000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Standard immunosuppressive treatment reduces regulatory B cells in children with autoimmune liver disease"],"prefix":"10.3389","volume":"13","author":[{"given":"Muhammed","family":"Yuksel","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Farinaz","family":"Nazmi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dima","family":"Wardat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sebahat","family":"Akg\u00fcl","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esra","family":"Polat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Murat","family":"Akyildiz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00c7igdem","family":"Arikan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2023,1,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.qjmed.a067264","article-title":"Controlled prospective trial of corticosteroid therapy in active chronic hepatitis","volume":"40","author":"Cook","year":"1971","journal-title":"Q J Med"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12843","article-title":"The induction of autoimmune hepatitis in the human leucocyte antigen-DR4 non-obese diabetic mice autoimmune hepatitis mouse model","volume":"186","author":"Yuksel","year":"2016","journal-title":"Clin Exp Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1002\/hep4.1202","article-title":"Immune alterations in patients with type 1 autoimmune hepatitis persist upon standard immunosuppressive treatment","volume":"2","author":"Renand","year":"2018","journal-title":"Hepatol Commun"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1097\/MPG.0000000000001801","article-title":"Diagnosis and management of pediatric autoimmune liver disease: ESPGHAN hepatology committee position statement","volume":"66","author":"Mieli-Vergani","year":"2018","journal-title":"J Pediatr Gastroenterol Nutr"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1002\/hep.31893","article-title":"Hla profile predicts severity of autoimmune liver disease in children of European ancestry","volume":"74","author":"Ma","year":"2021","journal-title":"Hepatology"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2013.08.004","article-title":"Autoimmune hepatitis: Contrasts and comparisons in children and adults - a comprehensive review","volume":"46","author":"Floreani","year":"2013","journal-title":"J Autoimmun"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1002\/hep.27998","article-title":"A novel \u201chumanized mouse\u201d model for autoimmune hepatitis and the association of gut microbiota with liver inflammation","volume":"62","author":"Yuksel","year":"2015","journal-title":"Hepatology"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1002\/hep.27884","article-title":"In autoimmune hepatitis type 1 or the autoimmune hepatitis-sclerosing cholangitis variant defective regulatory T-cell responsiveness to IL-2 results in low IL-10 production and impaired suppression","volume":"62","author":"Liberal","year":"2015","journal-title":"Hepatology"},{"key":"B9","doi-asserted-by":"publisher","first-page":"18017","DOI":"10.1038\/nrdp.2018.17","article-title":"Autoimmune hepatitis","volume":"4","author":"Mieli-Vergani","year":"2018","journal-title":"Nat Rev Dis Primers"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0181107","article-title":"Pediatric autoimmune hepatitis shows a disproportionate decline of regulatory T cells in the liver and of IL-2 in the blood of patients undergoing therapy","volume":"12","author":"Diestelhorst","year":"2017","journal-title":"PloS One"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/j.intimp.2016.05.014","article-title":"The role of regulatory b cells (Bregs) in the tregs-amplifying effect of sirolimus","volume":"38","author":"Song","year":"2016","journal-title":"Int Immunopharmacol"},{"key":"B12","doi-asserted-by":"publisher","first-page":"971","DOI":"10.1016\/j.jhep.2015.06.030","article-title":"EASL clinical practice guidelines: Autoimmune hepatitis","volume":"63","year":"2015","journal-title":"J Hepatol"},{"key":"B13","doi-asserted-by":"publisher","first-page":"IV1","DOI":"10.1136\/gut.45.2008.iv1","article-title":"Guidelines on the use of liver biopsy in clinical practice","author":"Grant","year":"1999","journal-title":"Br Soc Gastroenterology Gut"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1097\/MPG.0000000000003353","article-title":"Examining the hepatic immune system in children with liver disease with fine needle aspiration","volume":"74","author":"Yuksel","year":"2021","journal-title":"J Pediatr Gastroenterol Nutr"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1002\/hep.30475","article-title":"Normothermic machine perfusion (NMP) inhibits proinflammatory responses in the liver and promotes regeneration","volume":"70","author":"Jassem","year":"2018","journal-title":"Hepatology"},{"key":"B16","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1016\/j.immuni.2009.03.019","article-title":"Functional delineation and differentiation dynamics of human CD4+ T cells expressing the FoxP3 transcription factor","volume":"30","author":"Miyara","year":"2009","journal-title":"Immunity"},{"key":"B17","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1111\/cei.12956","article-title":"Immunosuppressive drugs affect interferon (IFN)-gamma and programmed cell death 1 (PD-1) kinetics in patients with newly diagnosed autoimmune hepatitis","volume":"189","author":"Grant","year":"2017","journal-title":"Clin Exp Immunol"},{"key":"B18","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1111\/ajt.16122","article-title":"Mucosal associated invariant T cells are differentially impaired in tolerant and immunosuppressed liver transplant recipients","volume":"21","author":"Sattler","year":"2021","journal-title":"Am J Transplant"},{"key":"B19","doi-asserted-by":"publisher","first-page":"525","DOI":"10.1038\/s41467-020-20874-x","article-title":"CTLA-4 expression by b-1a b cells is essential for immune tolerance","volume":"12","author":"Yang","year":"2021","journal-title":"Nat Commun"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2014.05.034","article-title":"Intrahepatic regulatory T cells in autoimmune hepatitis are associated with treatment response and depleted with current therapies","volume":"61","author":"Taubert","year":"2014","journal-title":"J Hepatol"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1002\/hep4.1163","article-title":"Changes in natural killer cells and exhausted memory regulatory T cells with corticosteroid therapy in acute autoimmune hepatitis","volume":"2","author":"Jeffery","year":"2018","journal-title":"Hepatol Commun"},{"key":"B22","doi-asserted-by":"publisher","first-page":"2671","DOI":"10.3390\/cells10102671","article-title":"Increased intrahepatic expression immune checkpoint molecules autoimmune liver disease","volume":"10","author":"Macek Jilkova","year":"2021","journal-title":"Cells"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1002\/hep.29782","article-title":"MAIT cells are chronically activated in patients with autoimmune liver disease and promote profibrogenic hepatic stellate cell activation","volume":"68","author":"Bottcher","year":"2018","journal-title":"Hepatology"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v27.i25.3705","article-title":"Incorporating mucosal-associated invariant T cells into the pathogenesis of chronic liver disease","volume":"27","author":"Czaja","year":"2021","journal-title":"World J Gastroenterol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1007\/s00281-022-00937-5","article-title":"B cells in autoimmune hepatitis: Bystanders or central players","volume":"44","author":"Schultheiss","year":"2022","journal-title":"Semin Immunopathol"},{"key":"B26","doi-asserted-by":"publisher","first-page":"411","DOI":"10.3390\/cells10020411","article-title":"Differential function of a novel population of the CD19+CD24hiCD38hi bregs in psoriasis and multiple myeloma","volume":"10","author":"Bartosinska","year":"2021","journal-title":"Cells"},{"key":"B27","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1186\/s12967-014-0251-9","article-title":"Increased numbers of CD5+CD19+CD1dhighIL-10+ bregs, CD4+Foxp3+ tregs, CD4+CXCR5+Foxp3+ follicular regulatory T (TFR) cells in CHB or CHC patients","volume":"12","author":"Wang","year":"2014","journal-title":"J Transl Med"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1103139","article-title":"IL-10-producing regulatory b cells in the pathogenesis of chronic hepatitis b virus infection","volume":"189","author":"Das","year":"2012","journal-title":"J Immunol"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1900488","article-title":"CD24(hi)CD38(hi) and CD24(hi)CD27(+) human regulatory b cells display common and distinct functional characteristics","volume":"203","author":"Hasan","year":"2019","journal-title":"J Immunol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1172\/JCI85113","article-title":"Human regulatory b cells in health and disease: Therapeutic potential","volume":"127","author":"Mauri","year":"2017","journal-title":"J Clin Invest"},{"key":"B31","doi-asserted-by":"publisher","first-page":"3019378","DOI":"10.1155\/2020\/3019378","article-title":"CD19(+)CD24(hi)CD38(hi) b cell dysfunction in primary biliary cholangitis","volume":"2020","author":"Chen","year":"2020","journal-title":"Mediators Inflamm"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1002\/hep.25682","article-title":"The impaired immune regulation of autoimmune hepatitis is linked to a defective galectin-9\/tim-3 pathway","volume":"56","author":"Liberal","year":"2012","journal-title":"Hepatology"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1097\/MEG.0000000000001804","article-title":"Immune monitoring of a child with autoimmune hepatitis and type 1 diabetes during COVID-19 infection","volume":"32","author":"Yuksel","year":"2020","journal-title":"Eur J Gastroenterol Hepatol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201948454","article-title":"Divergent chemokine receptor expression and the consequence for human IgG4 b cell responses","volume":"50","author":"Unger","year":"2020","journal-title":"Eur J Immunol"},{"key":"B35","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1084\/jem.20180595","article-title":"Immune regulation by glucocorticoids can be linked to cell type-dependent transcriptional responses","volume":"216","author":"Franco","year":"2019","journal-title":"J Exp Med"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1097\/00004836-200208000-00012","article-title":"Type 1 autoimmune hepatitis in Turkish patients: Absence of association with HLA B8","volume":"35","author":"Kosar","year":"2002","journal-title":"J Clin Gastroenterol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.4318\/tjg.2011.0155","article-title":"Relations between human leukocyte antigens and autoimmune hepatitis in Turkish children","volume":"22","author":"Geylani Gulec","year":"2011","journal-title":"Turk J Gastroenterol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2013.00442","article-title":"The repertoires of peptides presented by MHC-II in the thymus and in peripheral tissue: A clue for autoimmunity","volume":"4","author":"Collado","year":"2013","journal-title":"Front Immunol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1023\/A:1020166605016","article-title":"Genetic bases of autoimmune hepatitis","volume":"47","author":"Czaja","year":"2002","journal-title":"Dig Dis Sci"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840190311","article-title":"Allelic sequence variation in the HLA class II genes and proteins in patients with autoimmune hepatitis","volume":"19","author":"Doherty","year":"1994","journal-title":"Hepatology"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600-0528.2002.017401.x","article-title":"Genetic susceptibilities for immune expression and liver cell injury in autoimmune hepatitis","volume":"174","author":"Czaja","year":"2000","journal-title":"Immunol Rev"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1053216\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T11:46:48Z","timestamp":1672919208000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1053216\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,5]]},"references-count":41,"alternative-id":["10.3389\/fimmu.2022.1053216"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.1053216","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,5]]},"article-number":"1053216"}}