慢性HBV感染者外周血CD4^＋CD25^high调节性T细胞初步分析

Analysis of the CD4 ^＋ CD25^high regulatory T cells in peripheral blood of chronic hepatitis B virus infected patients

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^＋CD25^high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^＋CD25^highTreg含量，ELISA法检测HBsAg、HBsAb、HBeAg、HBeAb、HBcAb，RT—PCR法检测HBVDNA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／ul；（1．82±0．87）％）]及健康成人外周血标本[（8．91±3．11）个／ul，（1．35±0．39）％]中CD4^＋CD25^highTreg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^＋CD25^highTreg占CD4^＋T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^＋CD25^highTrge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^＋CD25^highTreg含量与临床指标间相关性发现，CD4^＋CD25^highTreg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HBVDNA水平无相关性（r=0．118，P〉0．05）；CD4^＋CD25^highTreg绝对计数与CD4／CD8比值呈正相关（r=0．360，P=0．015），与ALT水平及HBVDNA水平无相关性（r=-0．211，r=0．060，P〉0．05）。结论CD4^＋CD25^highTreg在HBV慢性感染的免疫发病机制中可能发挥一定作用。

Objective To analyze of CD4 ^＋ CD25^high 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^＋ CD25^highTreg 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^ ＋ CD25^high Treg and different clinical types of HBV infection and clinical indicators. Results The absolute counts of CD25^high Treg and its frequency in CD4 ^＋ T cells were similar between HBV infected patients [ （12.35 ±6.48）/ul, （1.82 ±0.87）% ] and health controls[ （8.91 ±3.11）/ul, （1.35 ±0.39）% ], P 〉 0.05. The frequency of CD25^high Treg in CD4^＋ T eeUs 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 CD25^high Treg from the immune tolerance group were significantly higher than the healthy control group（ P 〈 0.05 ）, and the frequency of CD25^highTreg 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 CD25^high 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^＋ CD25^high Treg may play a role in immunopatho- genesis of chronic HBV infection.