慢性乙肝患者外周血CD4^+CD25^+FOXP3^+Tregs及HBV抗原特异性CTLs的检测和分析

CD4^+CD25^+FOXP3^+ Tregs and HBV-specific CTLs in peripheral blood from chronic hepatitis B patients

目的:检测慢性乙肝(CHB)患者外周血中CD4+CD25+FOXP3+调节性T淋巴细胞(Treg细胞)和乙肝病毒(HBV)特异性细胞毒性T淋巴细胞(CTLs)的表达及意义。方法:收集28例CHB患者和15例健康人外周血单个核细胞标本,运用流式细胞仪对Treg细胞亚群进行定量分析,同时采用酶联免疫斑点法检测HBV抗原特异性CTLs,并结合丙氨酸氨基转移酶(ALT)和HBV DNA的临床情况进行分析。结果:CHB组CD4+CD25+FOXP3+Treg细胞的频率显著高于健康对照组(3.14%±0.97%vs 1.95%±0.68%,P<0.05);HBV抗原特异性CTL斑点计数为阳性(19.28±3.85)。CHB组Treg的频率与乙肝病毒载量呈正相关(r=0.831,P<0.01),与HBV特异性CTL斑点计数值呈负相关(r=-0.540,P<0.01)。结论:CHB患者外周血CD4+CD25+FOXP3+Treg细胞表达升高并与病毒载量相关,而与HBV反应的CTLs数量呈负相关,提示Treg细胞可通过抑制细胞免疫反应影响病毒清除。

AIM： To investigate the roles of CD4＋CD25＋FOXP3＋ regulatory T cells（Tregs） and HBV-specific cytotoxic T-lymphocytes（CTLs） in peripheral blood from the patients with chronic hepatitis B（CHB）.METHODS： Peripheral blood mononuclear cells from 28 patients with CHB and 15 healthy controls were analyzed for Treg frequency using flow cytometry and for HBV-specific CTLs using enzyme-linked immunospot assay（ELISPOT）.The clinical data of HBV-infected patients were considered.RESULTS： The frequency of CD4＋CD25＋FOXP3＋Tregs was higher in the patients with CHB than that in the patients of healthy controls（3.14%±0.97% vs 1.95%±0.68%,P0.05）,and a positive correlation was found between Tregs and the DNA levels of HBV（r=0.831,P0.01）.HBV-specific CTLs were detected by ELISPOT in CHB patients and a negative correlation was observed between Tregs and CTLs（r=-0.540,P0.01）.CONCLUSION： Peripheral blood CD4＋CD25＋FOXP3＋ Tregs in CHB patients are increased and closely correlated with the DNA replication of HBV and CTLs,suggesting that the clearance of HBV can be influenced by the inhibition of cellular immunoreaction through Tregs.