中文摘要：

目的采用T细胞表位预测软件结合体外实验鉴定丙型肝炎病毒（HCV）特异性细胞毒性T细胞（CTL）表位。方法采用T表位预测软件Rankpep预测HCV特异性CTL表位，选择候选CTL表位加以合成；用候选CTL表位肽分别刺激HCV感染者以及健康志愿者的外周血单个核细胞（PBMC），采用酶联免疫斑点试验（ELISPOT）检测PBMC中肽特异性分泌IFN-γ的斑点形成细胞（spots forming cells，SFC）的水平，采用细胞内细胞因子染色（intracellularcytokinestaining，ICS）检测PBMC中肽特异性IFN-γ＋CD8＋T细胞的水平。结果用5条候选CTL表位肽[NS3450（TVPQ—DAVSR）、NS3594（GPTPLLYRL）、NS4b78（SMMAFSAAL）、NS5a416（SEENVSVVF）和NS5a367（TVSSALAEL）]分别刺激10个HCV感染者和2个健康者的PBMC后，健康者的PBMC不产生IFN-γ，而7个HCV感染者的PBMC产生IFN-γ；HCV感染者的PBMC中肽特异性分泌IFN-γ的细胞的频率为（5～36）SFC／10^5PBMC，肽特异性IFN-γ＋CD8＋T细胞占总CD8＋T细胞的百分比为0．02％～0．25％。结论EL／SPOT结果和ICS结果证实5条肽NS3450、NS3594、NS4b78、NS5a416和NS5a367为全新的HCV特异性CTL表位。

英文摘要：

Objective To identify hepatitis C virus （HCV）-specific cytotoxicity T lymphocytes （CTL） epitopes by the combination of T epitopes prediction software and in vitro assays. Methods HCV- specific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-spe- cific CTL epitopes were selected. Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers, and then enzyme-linked immunospot （ELISPOT） and intra- cellular cytokine staining （ ICS ） were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ CD8 ＋ T cells in total CD8 ＋ T cells, respectively. Results Five can- didate CTL epitopes E NS3 450 （ TVPQDAVSR）, NS3 594 （ GPTPLLYRL）, NS4b 78 （ SMMAFSAAL）, NSSa 416（SEENVSVVF） and NSSa 367（TVSSALAEL）] were used to stimulate PBMC of ten HCV-infected pa- tients and two healthy volunteers. PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ. The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ＋ CD8 ＋ T cells ranged from 0.02% -0.25%. Conclusion Results of ELISPOT assay and ICS assay confirm that these five peptides NS3 450, NS3 594, NS4b 78, NS5a 416 and NSSa 367 are identified as novel HCV-specific CTL epitopes.