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Biliary atresia (BA) is a destructive pediatric liver disease and CD4 + T cell activation is demonstrated to play an important role in BA. However, a comprehensive scenario regarding the involvement of CD4 + T cell subsets to the development of BA remains unclear. Here, we aim to explore the infiltration of CD4 + T cell subsets and their clinical significance in BA. In the present study, thirty BA liver samples were collected during surgery and were divided into good (BA1,  n  = 16) and poor prognosis (BA2,  n  = 14), with samples from choledochal cyst patients ( n  = 8) as control. By using multiplex immunohistochemistry, we evaluated the infiltration level of CD4 + T cell subsets in the portal areas. RT-qPCR and flow cytometry were further applied to explore detailed features of Treg subsets. We revealed that hepatic infiltrating Th1, Th2, Th17, and ICOS + Treg cells were significantly increased in BA patients compared to controls and were negatively associated with prognosis, while high infiltrating ICOS − Tregs showed a favorable outcome. Phenotypic analysis indicated that, in contrast to ICOS + Tregs, ICOS − Tregs were mainly CD45RA hi CD45RO low , and preferentially expressed more CD73. Besides, RT-qPCR revealed elevated expression of  CD25, CD73, TGF- β, and  BCL-2  genes in ICOS − Tregs. Finally, functional assay confirmed that ICOS − Tregs had a higher suppressive capacity to cytokine secretion and were more resistant to apoptosis  in vitro . Collectively, we demonstrate that a mixed immune response is involved in BA pathogenesis, and the globally enhanced effector CD4 + T cell response is associated with unfavorable prognosis, highly suppressive ICOS − Tregs is a protective factor and may serve an important reference to predict prognosis.

CD4+T Cell Subset Profiling in Biliary Atresia Reveals ICOS− Regulatory T Cells as a Favorable Prognostic Factor