Bertoletti A, Ferrari C, Fiaccadori F, Penna A, Margolskee R, Schlicht H J, Fowler P, Guilhot S, Chisari F V
Department of Molecular and Experimental Medicine, Research Institute of Scripps Clinic, La Jolla, CA 92037.
Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10445-9. doi: 10.1073/pnas.88.23.10445.
Knowledge of the immune effector mechanisms responsible for clearance of hepatitis B virus (HBV)-infected cells has been severely limited by the absence of reproducible systems to selectively expand and to characterize HBV-specific cytotoxic T lymphocytes (CTLs) in the peripheral blood of patients with viral hepatitis. By using a strategy involving sequential stimulation with HBV nucleocapsid synthetic peptides followed by autologous, or HLA class I-matched, HBV nucleocapsid transfectants, we now report the existence of CTLs able to lyse target cells that express endogenously synthesized HBV nucleocapsid antigen in the peripheral blood of patients with acute viral hepatitis B. The CTL response is HLA-A2 restricted, mediated by CD8-positive T cells, and specific for a single epitope, located between amino acid residues 11 and 27 of HBV core protein; these residues are shared with the secretable precore-derived hepatitis B e antigen. Equivalent lysis of target cells that express each of these proteins suggests that their intracellular trafficking pathways may intersect. The current report provides definitive evidence that HLA class I-restricted, CD8-positive CTLs that recognize endogenously synthesized HBV nucleocapsid antigen are induced during acute HBV infection in humans and establishes a strategy that should permit a detailed analysis of the role played by HBV-specific CTLs in the immunopathogenesis of viral hepatitis.
由于缺乏可重复的系统来选择性扩增和鉴定病毒性肝炎患者外周血中乙肝病毒（HBV）特异性细胞毒性T淋巴细胞（CTL），对于负责清除HBV感染细胞的免疫效应机制的了解一直极为有限。通过采用一种策略，即先用HBV核衣壳合成肽进行序贯刺激，然后用自体或HLA I类匹配的HBV核衣壳转染细胞进行刺激，我们现在报告在急性乙型病毒性肝炎患者的外周血中存在能够裂解表达内源性合成HBV核衣壳抗原的靶细胞的CTL。该CTL反应受HLA - A2限制，由CD8阳性T细胞介导，且对位于HBV核心蛋白氨基酸残基11至27之间的单个表位具有特异性；这些残基与可分泌的前核心衍生的乙肝e抗原共有。对表达这些蛋白质中每一种的靶细胞的等效裂解表明它们的细胞内运输途径可能相交。本报告提供了确凿证据，表明在人类急性HBV感染期间可诱导识别内源性合成HBV核衣壳抗原的HLA I类限制的CD8阳性CTL，并建立了一种策略，该策略应能详细分析HBV特异性CTL在病毒性肝炎免疫发病机制中所起的作用。
