慢性HBV感染者外周血CD4+CD25high调节性T细胞初步分析

目的 分析不同临床乙型肝炎病毒(HBV)慢性感染者外周血中CD4+CD25high调节性T细胞(Treg)的水平及其与各种临床指标的关系.方法 采集35例不同临床表现成年慢性HBV感染者(HBsAb+组5例、非活动肝炎组8例、活动肝炎组12例、免疫耐受期组10例)及12例健康成人外周血标本,流式细胞仪分析外周血中CD4+CD25high Treg含量,ELISA法检测HBsAg、HBsAb、HBeAg、HBeAb、HBcAb,RT-PCR法检测HBV DNA载量,同时进行肝脏生化功能检测,并进行统计学分析.结果 HBV慢性感染者[(12.35±6.48)个/μl;(1.82 4-0.87)%)]及健康成人外周血标本[(8.91±3.11)个/μl,(1.35±0.39)%]中CD4+CD25highTreg绝对计数和其占CD4+T细胞百分含量差异均无统计学意义(P＞0.05);分组分析发现,免疫耐受期组CD4+CD25highTreg占CD4T细胞百分含量高于HBsAb+组、活动肝炎组及健康对照组(P＜0.05);免疫耐受期组CD4+CD25highTreg绝对计数高于健康对照组(P＜0.05);余各组间差异无统计学意义(P＞0.05).分析CD4+CD25highTreg含量与临床指标间相关性发现,CD4+CD25highTreg占CD4+T细胞百分含量与丙氨酸氨基转移酶(ALT)水平呈负相关(r=-0.418,P=0.038),与CD4/CD8比值呈正相关(r=0.344,P=0.021),与HBV DNA水平无相关性(r=0.118,P＞0.05);CD4+CD25highTreg绝对计数与CD4/CD8比值呈正相关(r=0.360,P=0.015),与ALT水平及HBV DNA水平无相关性(r=-0.211,r=-0.060,P＞0.05).结论 CD4+CD25highTreg在HBV慢性感染的免疫发病机制中可能发挥一定作用.

Analysis of the CD4+ CD25high regulatory T cells in peripheral blood of chronic hepatitis B virus in fected patients

Objective To analyze of CD4+ CD25high regulatory T cells(Treg)in peripheral blood of chronic HBV patients and its correlation with multiple clinical indicators.Methods Thirty-five hepatitis B virus(HBV)infected patients in this study were divided into four different clinical types:HBsAb+group(n=5),inactive hepatitis group(n=8),chronic hepatitis group(n=12),and immune tolerance group(n=10).The number of CD4+CD25high Treg and related T cells subgroup in CD3/CD4/CD8 was thoroughly examined by flow cytometry in peripheral blood of HBV infected patients and the healthy contrast group(n=12).Serum HBV markers were determined by commercial ELISA kits.Serum HBV DNA was quantified by commercial real-time PCR kit.Statistical differences were studied to investigate the correlations between CD4+CD25high Treg and different clinical types of HBV infection and clinical indicators.Results The absolute counts of CD25high Treg and its frequency in CD4+ T cells were similar between HBV infected patients [(12.35±6.48)/μ,(1.82±0.87)%]and health controls[(8.91±3.11)/μl,(1.35±0.39)%],P＞0.05.The frequency of CD25high Treg in CD4+ T cells from the immune tolerance group was significantly higher than that of the HBsAb+ group,chronic hepatitis group,and the healthy contrast group(P＜0.05).The absolute counts of CD25high Treg from the immune tolerance group were significantly higher than the healthy control group(P＜0.05),and the frequency of CD25high Treg in CD4+ T cells is negatively correlated to the ALT level(r=-0.418,P=0.038),positively correlated to CD4/CD8 ratio(r=0.344,P=0.021),no correlation to the HBV DNA level(r=0.118,P＞0.05).The absolute counts of CD25high Treg were positively correlated to CD4/CD8 ratio(r=0.360,P=0.015),no correlation to ALT level and HBV DNA level(r=-0.211,r=0.060,P＞0.05).Conclusion CD4+ CD25high Treg may play a role in immunopathogenesis of chronic HBV infection.