HLA-A2限制性NY-ESO-1b肽段诱导的CD8^+T细胞对稳定表达NY-ESO-1的HepG2肝癌细胞系的杀伤效应

Estimation of an NY-ESO-1 expressing HCC cell line by NY-ESO-1b specific CD8^+T cells in vitro induced by HLA-A2 restricted NY-ESO-1b peptide

目的分析体外诱导的NY-ESO-1特异性杀伤性T淋巴细胞对稳定表达NY-ESO-1蛋白的肝细胞性肝癌(hepatocellular carcinoma,HCC)细胞系的杀伤效应,为进一步分析NY-ESO-1蛋白作为疫苗用于治疗HCC的可能性提供实验支持。方法从HLA-A2阳性的肝癌患者体内分离出外周血单个核细胞,用NY-ESO-1肽段体外诱导特异性杀伤性T淋巴细胞,并通过酶联免疫斑点法分析特异性杀伤性T淋巴细胞对稳定表达NY-ESO-1蛋白的HCC细胞系HepG2细胞的杀伤效应。结果转染了NY-ESO-1的HepG2细胞(HLA-A2+)可有效地被NY-ESO-1157-165抗原肽诱导的特异性CD8+T淋巴细胞所识别。效应细胞中大量GrB的释放表明其对稳定转染了NY-ESO-1基因的HepG2细胞有很强的杀伤作用。结论HepG2细胞中表达的NY-ESO-1蛋白可被此肝癌细胞有效地加工处理,而且HLA-A2限制性的抗原肽NY-ESO-1157-165也可被有效地提呈到细胞表面,从而被效应T细胞所识别。

Objective： To analyze the effect of an NY-ESO-1 expressing HCC cell line by NY-ESO-lb specific CD8 ^＋ T cells in vitro induced by HLA-A2 Restricted NY-ESO-1 b peptide and to further analyze the possibility of NY-ESO-1 as a vaccine in HCC treatment. Methods： Peripheral blood mononuclear cells （PBMCs） were obtained from an HLA-A2^＋ HCC patient and NY-ESO-1 specific CD8 ^＋ T cells were in vitro induced by HLA-A2 Restricted NY-ESO-1 b peptide. Next we estimated the effect that NY-ESO-1 specific CD8 ^＋ T cells recognized and killed NY-ESO-1 expressing HepG2 ceils by enzyme-linked immunospot （ELISPOT）. Results： These in vitro induced NY-ESO-lb specific CD8 ^＋ T cells could recognize HepG2 cells stably transfected with NY-ESO-1 in both IFN-γ and Granzyme B ELISPOT assays. And the data also showed that NY-ESO-1 specific CD8 ~ T cells could efficiently kill NY-ESO-1 expressing HepG2 cells. Conclusion： NY-ESO-1 protein can be processed by HCC cells, and that HLA-A2 restricted NY- ESO-lb peptide （157-165） can also be presented on the surfaces of cells and recognized by NY-ESO-lb specific CD8 ^＋ T cells in vitro induced.