慢性乙型肝炎患者树突状细胞诱导的HBV特异性细胞毒性T淋巴细胞PD-1的表达  

Programmed cell death 1 expression on HBV-specific CTLs induced by dendritic cells in chronic hepatitis B patients

作　　者：彭建平[1] 孙克伟[1] 伍玉南[1] 张茜茜[1] 

机构地区：[1]湖南中医药大学第一附属医院传染科,长沙410007

出　　处：《临床肝胆病杂志》2012年第12期926-929,共4页Journal of Clinical Hepatology

基　　金：国家自然科学基金资助项目(No.81072779)

摘　　要：目的探讨慢性乙型肝炎(CHB)患者树突状细胞(DC)诱导的HBV特异性细胞毒性T细胞(CTL)表面程序性死亡受体1(PD-1)的表达情况及其与HBV DNA的关系。方法采集30例CHB患者和10例健康人的抗凝外周静脉血,分离外周血单个核细胞(PBMC),在白细胞介素(IL)-4和粒-巨噬细胞集落刺激因子(GM-CSF)的作用下培养使DC增殖、成熟,培养第4 d加入纯化的HBsAg进行冲击。采同一患者外周血,分离出自体T淋巴细胞,用含重组人白细胞介素(rhIL)-2的培养基维持T细胞的生长,培养第5 d与HBsAg冲击的DC共培养。以流式细胞技术检测CTL的PD-1表达,并分析PD-1表达水平与HBV DNA的关系。结果与健康对照组比较,CHB组DC诱导的HBV特异CTL的PD-1的表达明显升高(P=0.000)。且HBeAg阳性组PD-1的表达率较HBeAg阴性组明显升高(P=0.000)。CHB患者DC诱导的HBV特异性CTL的PD-1表达率与血清HBV DNA拷贝数的对数值呈正相关(r=0.53,P=0.008)。结论 CHB患者DC诱导的HBV特异性CTL高表达PD-1分子,为HBV慢性感染过程中CTL功能低下,病毒难以清除提供了另一条重要线索。Objective To investigate the expression of programmed cell death 1（PD - 1） surface receptor on hepatitis B virus（HBV） - specific cytotoxic T lymphocytes（CTLs） induced by dendritic cells（DCs） from patients with chronic hepatitis B（CHB）.Methods Peripheral blood mononuclear cells（PBMCs） were isolated from 30 CHB patients and 10 healthy controls.On the fourth day after inducement of differentiation into DCs in culture,the cells were pulsed with hepatitis B surface antigen（HBsAg） to induce HBV -specific DCs.The HBV -specificDCs were then co - cultured with autologous T cells isolated from homogeneous patients.The expression of PD - 1 was detected by flow cytometry. The relation between PD - 1 expression and HBV DNA load was assessed by Pearson＇s correlation analysis.Results The PD - 1 expression on HBV - specific CTLs induced by DCs was significantly higher in CHB patients（vs.healthy controls,P =0.000） and in CHB patients with positivity for hepatitis B e antigen（HBeAg）（vs.HBeAg - negative,P = 0.000）.In CHB patients,the PD - 1 expression level was positively correlated with serum HBV DNA load（r=0.53,P= 0.008）.Conclusion PD -1 expression is enhanced on HBV -specific CTLs induced by DCs from chronic hepatitis B patients.The up - regulated PD - 1 may contribute to the CTL dysfunction during HBV infection that is believed to promote survival and persistence of HBV.

关 键 词：肝炎病毒 乙型 肝炎 乙型 慢性 树突细胞 T淋巴细胞 细胞毒性 

分 类 号：R512.62[医药卫生—内科学]