肝癌患者外周血中针对NY-ESO-1/LAGE-1表位的自发性CTL的定量检测

Quantitative Detection of spontaneous NY-ESO-1-specific cytotoxic T lymphocyte in patients with hepatocellular carcinoma

目的检测原发性肝细胞癌(HCC)患者外周血中针对HLA-A2限制性NY-ESO-1b表位的特异性细胞毒性T细胞(cytotoxic Tlymphocyte,CTL)的水平,评价该表位多肽诱导HCC患者自发性细胞免疫应答的能力。方法采用流式细胞技术分析HCC患者的HLA表型,并通过RT-PCR技术检测其癌组织中NY-ESO-1/LAGE-1基因的表达情况;采用HLA-A2/多肽五聚体复合物即pentramer对HCC患者外周血中NY-ESO-1b表位特异性CTL进行标记并用流式细胞仪进行定量检测。结果在HLA-A2阳性、且其癌组织中NY-ESO-1/LAGE-1mRNA为阳性29例HCC患者中,13例(41.4%)患者外周血中可检测到针对HLA-A2限制性NY-ESO-1b表位(p157-165,SLLMWITQC)的特异性CTL应答,而在19例HLA-A2阳性的健康志愿者和12例HLA-A2阳性的肝炎后肝硬化患者外周血中均未检测到相应的自发性CTL应答(P值分别为0.004和0.023)。结论NY-ESO-1b表位多肽可诱发HCC患者体内产生较高频率的自发性CTL应答,提示该多肽可作为备选疫苗用于HCC的主动免疫治疗。

Objective To detect the frequency of the specific cytotoxic T lymphocytes（CTL）against HLA-A2 restricted NY-ESO- 1b epitope in the peripheral blood of patients with hepatocellular carcinoma （HCC） ,and evaluate the capacity of this epitope to induce spontaneous cellular immune response in HCC patients. Methods The HLA phenotypes of HCC patients were analysed by flow cytometry, and the expression of NY-ESO-1/LAGE-1 gene in their cancer tissues was detected by RT-PCR. The specific CTL against NY-ESO-1b epitope in the peripheral blood of HCC patients was marked using HIA-A2 pentameric complexes and quantified byflow cytometry.Results Out of the 29 NY-ESO-1 mRNA＋ HLA-A2＋ HCC patients the specific CD8＋ T-cell responses to NY：ESO-lb epitope（p157-165, SLLMWITQC ）could be detected in 13 patients（ 41.4 % ）;while the spontaneous CTL were not detected in the peripheral blood of19 HLA-A2＋ healthy volunteers（p value was 0.004） and 12 HEA-A2＋ cirrhotic patients due to hepatitis（p value was 0.023）. Conclusion NY-ESO-lb epitope can induce spontaneous CTL responses with a relatively high frequency in HCC patients, which suggest that this peptide can be used as a vaccine candidate for active immunotherapy of HCC.