乙型肝炎病毒感染不同阶段患者外周血CD4^＋CD25^＋调节性T细胞的测定  

Study on circulating CD4^＋ CD25^＋ regulatory T cells in different stages of hepatitis B virus infected patients

作　　者：彭国平[1] 孙雯[2] 孙箴[1] 蔡乃青[2] 陈智[1] 

机构地区：[1]浙江大学医学院附属第一医院传染病研究所,传染病诊治国家重点实验室,杭州310003 [2]浙江大学医学院附属口腔医院正畸科

出　　处：《中华传染病杂志》2008年第1期40-45,共6页Chinese Journal of Infectious Diseases

摘　　要：目的测定HBV感染不同阶段患者外周血CD4^＋CD25^＋调节性T细胞（Treg）的频率及标记分子，并分析其与临床指标的相关性。方法采集79例慢性乙型肝炎（CHB）、12例急性乙型肝炎（AHB）患者、26例无症状HBV携带者（ASC）和20例健康对照的外周血，流式细胞仪分析Treg频率、Treg细胞表面和胞内特征性分子的表达。普通RT—PCR和相对荧光定量PCR测定叉头／翼状转录因子3（Foxp3）在CD25^＋Treg细胞的表达水平。所有患者及健康对照均经ELISA检测HBV血清标记物水平，实时荧光定量PCR测定血清HBVDNA载量，并进行肝功能检测。结果总CD4^＋CD25^＋T细胞占外周血CD4^＋T细胞的比率，在各患者组和健康对照组之间差异均无统计学意义（P〉0．05）。HBeAg阳性CHB组CD4^＋CD25高表达T细胞频率（3．42％±0．81％）与HBeAg阴性CHB组（3．19％±0．67％）、ASC组（3．05％±0．64%）比较，差异无统计学意义（均P〉0．05），但明显高于健康对照组（2．72％±0．71％，P=0．034）和AHB组（2．25％±0．54％，P=0．013）。CD4^＋CD25高表达T细胞表面高表达CD45RO、CD25分子，低表达CD45RA，细胞内高表达细胞毒性T淋巴细胞相关抗原-4（CTLA-4）和Foxp3。各患者组及健康对照组Treg中Foxp3表达水平的差异无统计学意义（P〉0．05）。CHB组患者的Treg频率与血清病毒载量呈正相关（r=0．48，P=0．018）。结论Treg可能通过抑制T细胞免疫应答反应而影响病毒清除，并与CHB患者的持续感染密切相关。Objective To analyze the frequency and characteristics of circulating CD4^＋ CD25^＋ regulatory T cells（Treg） in different stages of hepatitis B virus（HBV）-infected patients, and to inves- tigate the correlation between circulating Treg and serum HBV DNA loads. Methods Seventy-nine chronic hepatitis B（CHB） patients, 12 acute hepatitis B（AHB） patients, 26 asymptomatic HBV carriers（ASC） and 20 healthy blood donators were enrolled in this study. The frequency and characteristics of Treg in peripheral blood from different subjects were analyzed using flow cytometry. The Forkhead box P3（Foxp3） expression level in Treg was detected by regular reverse transcriptase-polymerase chain reaction（RT-PCR） and quantitative real-time PCR. Serum HBV titers were measured using real-time PCR, and HBV markers and liver function were also evaluated for each subject. Results HBeAg positive CHB patients presented a higher fraction of circulating CD25 high Treg （3.42% ± 0.81% ） than those in AHB patients（2.25% ± 0.54%, P = 0. 013） and healthy controls （2.72%± 0. 71%, P = 0. 034）, but were similar to HBeAg negative CHB patients（3. 19% ± 0.67 % ） and ASC patients（3.05 % ± 0.64 % ）. And there was no significant difference of total CD4^＋ CD25^＋ T cell frequency among all tested groups. There was no significant difference of Foxp3 expression in Treg from different groups. The CD4^＋ CD25 high T subset had a high surface-expression of CD45RO and intracellular-expression of cytotoxic T lymphocyte related antigen-4 （CTLA-4） and Foxp3. Furthermore, the frequency of circulating Treg in CHB patients was positively associated with HBV DNA level（r=0. 48, P= 0. 018）, but not with serum alanine aminotransferase level（P〉0.05）. Conclusions The CD4^＋ CD25^＋ Treg could probably influence the clearance of virus by suppressing the antiviral T cell responses in CHB patients, and the increased circulating Treg is positively associated with HBV DNA titers.

关 键 词：肝炎 乙型 CD4阳性T淋巴细胞 受体 白细胞介素2 免疫耐受 FOXP3 

分 类 号：R51[医药卫生—内科学]