Aberrant expression of activation regulators CBL-B, CTLA-4 and PD-1 in intrahepatic Teff cells in autoimmune hepatitis
Aberrant expression of activation regulators CBL-B, CTLA-4 and PD-1 in intrahepatic Teff cells in autoimmune hepatitis
复制标题
自身免疫性肝炎肝内Teff细胞激活调节因子CBL-B、CTLA-4和PD-1的异常表达
DOI：
10.1055/s-0039-3402250
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
M Sebode
中科院分区：
文献类型：
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作者：
P Filpe;A Wöstemeier;E De Martin;S Weidemann;R Taubert;C Schramm;AW Lohse;J Herkel;M Sebode
方法：对86例AIH患者进行检查。通过定量聚合酶链式反应(QPCR)检测AIH患者和正常对照组外周血T细胞活化调节因子CBL-B、CTLA-4、GRAIL、ICOS、ITCH、NEDD4、OX40、PD-1、PKC((Theta))和TRAF6的基因表达水平。对AIH和DILI患者的肝组织标本进行RNA原位杂交和CD3联合染色。肝组织学采用改良肝活动指数(MHAI)评分，定量评价肝内炎症活动程度。结果：定量聚合酶链式反应显示初治AIH患者肝组织中Cbl-B(p<；0.001)、PD-1(p<；0.01)和CTLA-4(p<；0.001)均显著升高。AIH患者肝内CBL-B、PD-1和CTLA-4的表达与MHAI呈正相关。RNA原位杂交显示初治AIH患者肝门区CBL-B(p<；0.01)、CTLA-4(p<；0.01)和PD-1(p<；0.02)阳性T细胞明显高于DILI和治疗中AIH患者。流式细胞仪检测显示，初治AIH患者肝内Cbl-B+CD4+T细胞(p<；0.005)和Cbl-B+CD8+T细胞(p<；0.005)水平均高于健康对照组。结论：活动期AIH患者肝内T细胞活化后仍保持较高水平的激活抑制物CbL-B，这与以往在小鼠和多发性硬化患者中的研究结果不同。CBL-B的表达与AIH的组织学活动呈正相关。此外，活动期AIH患者肝内T细胞中CBL-B调节剂CTLA-4和PD-1的表达也上调。提示肝内T细胞活化调节因子在AIH中异常表达，参与了AIH的发病机制。
Methods:A total of 86 AIH patients were examined. Gene expression levels of activation regulators CBL-B, CTLA-4, GRAIL, ICOS, ITCH, NEDD4, OX40, PD-1, PKC ((Theta)) and TRAF6 were analysed by qPCR of liver samples or isolated peripheral T cells of AIH patients and control subjects. RNA in-situ hybridization with selected probes and CD3 co-staining was performed on liver samples of AIH or DILI patients. The modified hepatic activity index (mHAI) score was applied to liver histologies to quantify intrahepatic inflammatory activity. Protein expression of intrahepatic T effector cells was examined by flow cytometry.Results:qPCR screening revealed that in livers of treatment-naïve AIH patients as compared to healthy control subjects, CBL-B (p< 0.01), PD-1 (p< 0.01) and CTLA-4 (p< 0.001) were significantly elevated. Intrahepatic expression of CBL-B, PD-1 and CTLA-4 in AIH patients correlated positively with the mHAI. RNA in-situ hybridization revealed that T cells positive for CBL-B (p< 0.01), CTLA-4 (p< 0.01) and PD-1 (p< 0.02) were increased in hepatic portal areas of treatment-naïve AIH patients as compared to DILI patients and to AIH patients under treatment. Flow cytometry revealed that levels of intrahepatic CBL-B+ CD4+ T cells (p< 0.005) and CBL-B+ CD8+ T cells (p< 0.005) were elevated in treatment-naïve AIH patients as compared to healthy controls. CTLA-4+ CD4+ T cells (p< 0.005), CTLA-4+ CD8+ T cells and PD-1+ CD8+ T cells were increased in treatment-naïve AIH patients in comparison to healthy controls.Conclusion:Intrahepatic T cells in active AIH maintain high levels of the activation inhibitor CBL-B following activation, which contrasts with previous studies in mice and in patients with multiple sclerosis. CBL-B expression correlated positively with histological activity of AIH. Moreover, the CBL-B modulators CTLA-4 and PD-1 were also up-regulated in intrahepatic T cells in active AIH. These findings indicate that intrahepatic T cell activation regulators are aberrantly expressed in AIH, thus contributing to its pathogenesis.