Zhang Shuhao, Goswami Shyamal, Ma Jiaqiang, Meng Lu, Wang Youping, Zhu Fangming, Zhang Dandan, Zheng Shan, Dong Rui, Xiao Xianmin, Zhang Xiaoming, Chen Gong
Department of Pediatric Surgery, Children's Hospital of Fudan University, Shanghai, China.
Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
Front Pediatr. 2019 Jul 9;7:279. doi: 10.3389/fped.2019.00279. eCollection 2019.
Biliary atresia (BA) is a destructive pediatric liver disease and CD4T cell activation is demonstrated to play an important role in BA. However, a comprehensive scenario regarding the involvement of CD4T cell subsets to the development of BA remains unclear. Here, we aim to explore the infiltration of CD4T 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, = 16) and poor prognosis (BA2, = 14), with samples from choledochal cyst patients ( = 8) as control. By using multiplex immunohistochemistry, we evaluated the infiltration level of CD4T 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 ICOSTreg cells were significantly increased in BA patients compared to controls and were negatively associated with prognosis, while high infiltrating ICOSTregs showed a favorable outcome. Phenotypic analysis indicated that, in contrast to ICOSTregs, ICOSTregs were mainly CD45RACD45RO, and preferentially expressed more CD73. Besides, RT-qPCR revealed elevated expression of β, and genes in ICOSTregs. Finally, functional assay confirmed that ICOSTregs had a higher suppressive capacity to cytokine secretion and were more resistant to apoptosis . Collectively, we demonstrate that a mixed immune response is involved in BA pathogenesis, and the globally enhanced effector CD4T cell response is associated with unfavorable prognosis, highly suppressive ICOSTregs is a protective factor and may serve an important reference to predict prognosis.
胆道闭锁（BA）是一种具有破坏性的儿科肝脏疾病，已证实CD4T细胞激活在BA中起重要作用。然而，关于CD4T细胞亚群参与BA发展的全面情况仍不清楚。在此，我们旨在探讨CD4T细胞亚群在BA中的浸润情况及其临床意义。在本研究中，手术期间收集了30份BA肝脏样本，分为预后良好组（BA1，n = 16）和预后不良组（BA2，n = 14），并将胆总管囊肿患者的样本（n = 8）作为对照。通过多重免疫组化，我们评估了门管区CD4T细胞亚群的浸润水平。进一步应用RT-qPCR和流式细胞术来探究调节性T细胞（Treg）亚群的详细特征。我们发现，与对照组相比，BA患者肝脏中浸润的辅助性T细胞1（Th1）、辅助性T细胞2（Th2）、辅助性T细胞17（Th17）和诱导共刺激分子（ICOS）调节性T细胞显著增加，且与预后呈负相关，而高浸润的ICOS调节性T细胞显示出良好的预后。表型分析表明，与ICOS调节性T细胞相反，ICOS调节性T细胞主要为CD45RA⁺CD45RO⁻，并优先表达更多的CD73。此外，RT-qPCR显示ICOS调节性T细胞中β、γ和δ基因的表达升高。最后，功能分析证实ICOS调节性T细胞对细胞因子分泌具有更高的抑制能力，并且对凋亡更具抗性。总体而言，我们证明混合免疫反应参与BA发病机制，整体增强的效应性CD4T细胞反应与不良预后相关，高抑制性的ICOS调节性T细胞是一个保护因素，可能为预测预后提供重要参考。
