基于单细胞测序技术评估乙型肝炎相关慢加急性肝衰竭患者外周血T淋巴细胞亚群特点  

Evaluation of peripheral blood T-lymphocyte subpopulations features in patients with hepatitis B virus-related acute-on-chronic liver failure based on single-cell sequencing technology

作　　者：彭鹏[1] 季亚秋 赵凝慧 刘甜 王瀚 姚佳 Peng Peng;Ji Yaqiu;Zhao Ninghui;Liu Tian;Wang Han;Yao Jia(Department of Gastroenterology,Shanxi Provincial People's Hospital,Taiyuan 030031,China;Department of Biochemistry and Molecular Biology,Basic Medical College,Shanxi Medical University,Taiyuan 030000,China;Department of Gastroenterology,Shanxi Bethune Hospital,Taiyuan 030032,China)

机构地区：[1]山西省人民医院消化内科,太原030031 [2]山西医科大学基础医学院生物化学与分子生物学教研室,太原030000 [3]山西白求恩医院消化科,太原030032

出　　处：《中华肝脏病杂志》2023年第4期422-427,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金青年基金(81700562);山西省136兴医工程(普外科部分2021YZ13);山西省基础研究项目(202103021224341、202203021222342);山西省卫健委引导性科技专项(2021XM42)。

摘　　要：目的耗竭T细胞是免疫功能障碍的重要组成部分。探讨乙型肝炎病毒相关慢加急性肝衰竭(HBV-ACLF)患者外周血耗竭T淋巴细胞特点,为ACLF免疫功能障碍患者提供潜在的治疗靶点分子。方法选取6例HBV-ACLF患者和3例健康对照者采用单细胞RNA测序方法进行T细胞异质性检测,并且筛选耗竭T淋巴细胞亚群,分析其基因表达特点,并且通过拟时间分析其发展轨迹。两组样本之间比较采用独立样本t检验分析。结果HBV-ACLF患者外周血中T淋巴细胞具有不同的分化轨迹,可分化为8个具有不同特征的亚群,其中高度表达耗竭基因的CD4^(+)TIGIT^(+)亚群(P=0.007)和CD8^(+)LAG3^(+)(P=0.010)亚群明显高于健康对照者。拟时间分析显示,CD4^(+)TIGIT^(+)亚群和CD8^(+)LAG3^(+)亚群在T淋巴细胞分化的后期出现,提示T细胞在ACLF发病过程中从幼稚-效应-耗竭的转变。结论HBV-ACLF患者外周血T细胞分化存在异质性,以CD4^(+)TIGIT^(+)T细胞亚群和CD8^(+)LAG3^(+)T细胞亚群为特征的耗竭T细胞数目明显增加,提示T细胞的免疫耗竭参与HBV-ACLF的免疫功能障碍,为改善ACLF患者免疫功能障碍提供潜在有效的靶点分子。Objective T lymphocyte exhaustion is an important component of immune dysfunction.Therefore,exploring peripheral blood-exhausted T lymphocyte features in patients with hepatitis B virus-related acute-on-chronic liver failure may provide potential therapeutic target molecules for ACLF immune dysfunction.Methods Six cases with HBV-ACLF and three healthy controls were selected for T-cell heterogeneity detection using the single-cell RNA sequencing method.In addition,exhausted T lymphocyte subpopulations were screened to analyze their gene expression features,and their developmental trajectories quasi-timing.An independent sample t-test was used to compare the samples between the two groups.Results Peripheral blood T lymphocytes in HBV-ACLF patients had different differentiation trajectories with different features distinct into eight subpopulations.Among them,the CD4^(+)TIGIT^(+)subsets(P=0.007)and CD8^(+)LAG3^(+)(P=0.010)subsets with highly exhausted genes were significantly higher than those in healthy controls.Quasi-time analysis showed that CD4^(+)TIGIT^(+)and CD8^(+)LAG3^(+)subsets appeared in the late stage of T lymphocyte differentiation,suggesting the transition of T lymphocyte from naïve-effector-exhausted during ACLF pathogenesis.Conclusion There is heterogeneity in peripheral blood T lymphocyte differentiation in patients with HBV-ACLF,and the number of exhausted T cells featured by CD4^(+)TIGIT^(+)T cell and CD8^(+)LAG3^(+)T cell subsets increases significantly,suggesting that T lymphocyte immune exhaustion is involved in the immune dysfunction of HBV-ACLF,thereby identifying potential effective target molecules for improving ACLF patients'immune function.

关 键 词：慢加急性肝衰竭 T淋巴细胞 单细胞RNA测序 耗竭T细胞 

分 类 号：R512.62[医药卫生—内科学] R575.3[医药卫生—临床医学]