乙型肝炎病毒特异性细胞毒性T淋巴细胞应答与不同临床感染状态的关系  

The association between antigen-specific cytotoxic lymphocytes response and different clinical status in patients with hepatitis B

作　　者：万玉峰[1] 韩亚萍[1] 李军[1] 孔练花[1] 李爽[1] 董莉[1] 陈念[1] 刘源[1] 黄祖瑚[1] 

机构地区：[1]南京医科大学第一附属医院感染病科,210029

出　　处：《中华传染病杂志》2009年第5期287-291,共5页Chinese Journal of Infectious Diseases

基　　金：江苏省科教兴卫工程医学重点学科工程资助项目（200707）;江苏省卫生厅医学科技发展基金资助项目（H200311）;江苏省医学重点人才资金资助项目（RC2007049）

摘　　要：目的分析人类白细胞抗原（HLA）-A0201限制性的特异性CTL，研究急性肝炎急性期和慢性乙型肝炎活动期患者T淋巴细胞对特异性抗原表位免疫应答的差异。方法收集HLA—A0201阳性的5例急性肝炎急性期和6例慢性乙型肝炎活动期患者的外周血单个核细胞（PBMc），酶联免疫斑点技术（EI，ISPOT）测定针对HBV聚合酶区（P01575—583）、包膜区（Env348—357）和核心区（Core18—27）3个CD8^＋T淋巴细胞表位肽特异性CTL的数量和功能。数据采用t检验。结果经P01575583、Env348—357和Core18—27三条抗原肽刺激，急性乙型肝炎急性期患者组斑点形成细胞数（SFC）分别为110±13、165±17和185±20；慢性乙型肝炎活动期患者组SFC分别为22±4、23±5和30±5，两组差异有统计学意义（f值分别为10．9、15．2和8．0，均P〈0．05）。急性乙型肝炎急性期患者各抗原肽特异性CTL的应答能力P01575—583〈Env348—357〈Corel8-27，其中P01575—583和Core18-27比较，差异有统计学意义（t=4．0，P〈0．05），而Env348—357和Core18—27比较，差异无统计学意义（P〉0．05）。非特异性HLA-2402限制性Corell7-125刺激也出现SFC增加，但与阴性对照组比较，差异无统计学意义（P〉0．05）。结论急性感染者HBV特异性CTL应答水平显著高于慢性HBV感染者，慢性乙型肝炎患者体内的多克隆CTL数量和功能低下。Objective To analyze human leucocyte antigen （HLA）-A0201 restricted antigenspecific cytotoxic lymphocytes （CTL）, and to investigate the difference of T cell response to specific antigen epitopes between patients with acute phase of acute hepatitis B and active phase of chronic hepatitis B. Methods Peripheral blood mononuclear cells （PBMC） from 5 patients with acute phase of acute hepatitis B and 6 patients with active phase of chronic hepatitis B were isolated. The numbers and functions of CD8^＋ T lymphocyte epitope peptide specific CTL were detected using enzyme-linked immunosorbent spot （EI.ISPOT） assay, and the 3 peptides were from HBV polymerase region （Po1575-583）, envelope region （Env348-357） and core region （Core18-27）, respectively. The data were analyzed using t test. Results The spot formation cell counts （SFC） of Po1575-583, Env348-357 and Core18-27 stimulations in patients with acute phase of acute hepatitis B were 110±13, 165 ±17 and 185 ± 20, respectively; and those in patients with active phase of chronic hepatitis B were 22 ± 4,23±5 and 30± 5, respectively; the differences were all significant （t = 10.9, 15.2 and 8.0, respectively, all P〈0.05）. The CTL responses to the three peptides in patients with acute phase of acute hepatitis B were Po1575-583〈Env348 357〈Core18-27; and the difference between responses to Po1575-583 and Core18-27 was significant （t=4.0, P〈0. 05）, while there was no statistical difference between CTL responses to Env348-357 and Corel8 27 （P〉0.05）. The SFC were increased upon nonantigen specific HLA-A2404 restricted epitope （Core117-125）, but the difference was not significant compared with negative control group （P 〉 0. 05）. Conelusions Hepatitis B virus-specific CTL responses in patients with acute hepatitis B are significantly higher than those in patients with chronic hepatitis B. The number and function of polyclonal CTL are both impaired in patients with chronic hepatitis B.

关 键 词：T淋巴细胞 细胞毒性 HLA抗原 肝炎 乙型 酶联免疫斑点技术 肝炎病毒 乙型 

分 类 号：R686[医药卫生—骨科学]