急性期恶性疟患者外周血PD-1^(+)TIGIT^(+)T细胞功能  

Function of PD-1^(+)TIGIT^(+)T cells in peripheral blood of falciparum malaria patients in acute phase

作　　者：黄雅倩 王蓓蓓[1] 宋蕊[2] 徐晓雪 赵雪 王芯栎 陈晨[1] Huang Yaqian;Wang Beibei;Song Rui;Xu Xiaoxue;Zhao Xue;Wang Xinyue;Chen Chen(Institute of Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,100015 Beijing,China;Infectious Diseases Department-,Beijing Ditan Hospital,Capital Medical University,100015 Beijing,China)

机构地区：[1]首都医科大学附属北京地坛医院传染病研究所,北京100015 [2]首都医科大学附属北京地坛医院感染二科,北京100015

出　　处：《中华实验和临床感染病杂志（电子版）》2021年第6期385-393,共9页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金面上项目(No.81971862);北京市医院管理中心“登峰”计划专项经费资助(No.DFL20191801)。

摘　　要：目的研究急性期恶性疟患者外周血T淋巴细胞共抑制分子PD-1和TIGIT的表达模式及功能。方法收集2019年4月至2019年10月首都医科大学附属北京地坛医院感染二科14例非洲输入型急性期恶性疟患者发病后4~12 d和28例健康对照者外周血单个核细胞。流式细胞术检测患者外周血CD4^(+)T和CD8^(+)T淋巴细胞PD-1^(+)TIGIT^(-)、PD-1^(+)TIGIT^(-)、PD-1-TIGIT+和PD-1^(+)TIGIT^(+)四个亚群比例,比较各细胞亚群凋亡(Annexin V+)、增殖(Ki67+)、分泌细胞因子(IFN-γ^(+)、IL-2^(+)和TNF-α^(+))和杀伤(Granzyme B+和Perforin+)能力差异,正态分布资料采用one-way ANOVA检验进行方差分析,Holm-Sidak’s进行多重比较;偏态分布资料则采用Kruskal-Wallis检验进行分析,Bonferroni法进行多重比较。统计患者临床资料并分析急性期恶性疟患者T细胞各亚群比例和CD8^(+)T细胞杀伤能力与临床指标的相关性,正态分布资料采用Pearson相关性分析,非正态分布资料采用Spearman相关性分析。结果与健康对照组相比,急性期恶性疟患者CD4^(+)T和CD8^(+)T细胞中PD-1^(+)和PD-1^(+)TIGIT^(+)细胞比例均显著增高(CD4:t=4.85、P<0.001,t=4.77、P=0.0002;CD8:t=3.90、P=0.001、t=3.29、P=0.0021);CD4^(+)T细胞TIGIT+亚群增加(t=4.12、P=0.0002)。CD4^(+)T细胞和CD8^(+)T细胞PD-1^(+)TIGIT^(-)和PD-1^(+)TIGIT^(+)细胞亚群Annexin V+表达比例均显著高于PD-1^(+)TIGIT^(-)细胞亚群(F=4.67、P=0.0125,F=3.19、P=0.0460),但CD4^(+)T细胞PD-1^(+)TIGIT^(-)和PD-1^(+)TIGIT^(+)细胞亚群Ki67+和IFN-γ^(+)表达比例均显著高于PD-1^(+)TIGIT^(-)细胞亚群(H=17.80、P=0.0005,F=11.22、P=0.0002),CD8^(+)T细胞PD-1^(+)TIGIT^(-)和PD-1^(+)TIGIT^(+)细胞亚群IFN-γ^(+)、IL-2^(+)和Granzyme B+表达比例均显著高于PD-1^(+)TIGIT^(-)细胞亚群(F=7.95、P=0.047,H=8.29、P=0.0404,F=12.18、P<0.001)。急性期恶性疟患者CD4^(+)和CD8^(+)T细胞各亚群比例与患者临床参数相关性分析发现CD4^(+)T细胞PD-1^(+)TIGIT^(+)表达比Objective To investigate the expression patterns and potential functions of PD-1 and TIGIT,two co-inhibitory molecules,on T lymphocyte cells in peripheral blood of patients with acute falciparum malaria infection.Methods Fourteen African imported patients with acute falciparum malaria in Infectious Diseases Department-Ⅱof Beijing Ditan Hospital,Capital Medical University,from April to October 2019 were enrolled.Peripheral blood mononuclear cells at 4-12 d after symptom onset were collected,while 28 healthy were enrolled as controls.Flow cytometry was used to detect the proportions of four subsets(PD-1^(+)TIGIT^(-),PD-1^(+)TIGIT^(-),PD-1-TIGIT+and PD-1^(+)TIGIT^(+))of CD4^(+)T and CD8^(+)T cells in peripheral blood of patients,and the further expression level on apoptosis(Annexin V+),proliferation(Ki67+),cytokines secretion(IFN-γ^(+),IL-2^(+)and TNF-α^(+))and cytotoxicity(Granzyme B+and Perforin+)in four subsets were compared,among which,variance analysis of normal distribution data were analyzed by one-way ANOVA test and multiple comparison were taken by holm-Sidak’s test.Skewness distribution data were analyzed by Kruskal-wallis test for variance analysis andmultiple comparison were taken by Bonferroni method.The clinical data of patients were collected,and correlations between the proportion of T cell subsets and clinical parameters as well as the cytotoxicity of CD8^(+)T cells and clinical parameters in patients with acute falciparum malaria were analyzed,among which,normal distribution data were analyzed by Pearson correlation analysis and skewness distribution data were analyzed by Spearman correlation analysis.Results Compared with healthy controls,the proportions of PD-1^(+)and PD-1^(+)TIGIT^(+)subsets of CD4^(+)and CD8^(+)T cells in acute falciparum malaria patients were significantly increased(CD4:t=4.85,P<0.001;t=4.77,P=0.0002;CD8:t=3.90,P=0.001;t=3.29,P=0.0021).The proportion of TIGIT+subsets of CD4^(+)T cells was increased(t=4.12,P=0.0002).The rates of Annexin V+expressed by PD-1^(+)TIGIT^(-)and

关 键 词：恶性疟 急性期 T淋巴细胞 共抑制分子 多色流式细胞术 

分 类 号：R531.3[医药卫生—内科学]