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

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(ELISPOT)测定针对HBV聚合酶区(Pol575-583)、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Pol575-583、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(t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Pol575-5830.05).非特异性HLA-2402限制性Core117-125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.

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

Objective To analyze human leucocyte antigen (HLA)-A0201 restricted antigen-specific 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 (ELISPOT) assay, and the 3 peptides were from HBV polymerase region (Pol575-583), 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 Pol575-583, 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 Pol575-5830.05). The SFC were increased upon non-antigen specific HLA-A2404 restricted epitope (Core117-125), but the difference was not significant compared with negative control group (P>0.05). Conclusions 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.