HLA-A^＊0201/CAP-1四聚体在结直肠癌研究中的应用

Application of HLA-A^＊0201/CAP-1 tetramer in colorectal cancer

目的:应用HLA-A*0201/CAP-1四聚体,分析HLA-A*0201+CEA+结直肠癌患者的外周血及癌旁肠系膜引流淋巴结的特异性CTLs的数量。方法:分离健康人(18例)PBMC和结直肠癌(23例)患者PBMC以及癌旁肠系膜引流淋巴结的淋巴细胞,应用流式细胞术,以HLA-A*0201/CAP-1和HLA-A*0201/FLUmp四聚体检测CAP-1和FLUmp特异性CTLs的频率。结果:在25例HLA-A*0201+个体中,结直肠癌组和正常组外周血单个核细胞中HLA-A*0201/FLU四聚体+CD8+细胞的频率分别为(0.671±0.421)%和(0.564±0.408)%,两组之间无显著差异(P=0.525)。结直肠癌组和正常组外周血单个核细胞中HLA-A*0201/CAP-1四聚体+CD8+细胞的频率分别为(2.409±2.385)%和(0.020±0.021)%,两组之间有显著差异(P=0.008)。结论:HLA-A*0201+结直肠癌患者HLA-A*0201/CAP-1四聚体+CD8+细胞的频率升高,说明CEA特异性CTLs在结直肠癌患者的免疫监视功能中具有重要的作用。对存在针对肿瘤抗原的特异性CTLs却无法阻止肿瘤进展的具体机制需要更加深入的研究。

Objective： To analyze the frequencies of HLA-A^＊0201 restricted CEA-specific CD8＋T cells,HLA-A^＊0201 /FLUmp tetramer and HLA-A^＊0201 /CAP-1 tetramer were applied in patients with colorectal cancer. Methods： Lymphocytes from peripheral blood and lymph node,1 × 106 cells /ml,were incubated with 1 μg HLA-A^＊0201 /peptide tetramers and anti-CD8 for 1 h at 25 coseperately. The cells were then washed in PBS. Next,the cells were illuminated by detecting frequencies of FLUmp-specific CD8＋T cells and CAP-1-specific CD8＋T cells with flow cytometry. Results： HLA-A^＊0201 /peptide were used to detect CAP-1 or FLUmp-specific CD8＋T cells,which were analyzed either healthy individuals or patients with colorectal cancer. We did not find differences in average frequencies of FLUmp-specific CD8＋T cells between 11 HLA-A^＊0201＋patients with colorectal cancer and 14 HLA-A＊0201＋healthy individuals [（ 0. 671 ± 0. 421） %,（ 0. 564 ± 0. 408） %]. But the frequencies of CAP-1-specific CD8＋T cells of HLA-A^＊0201＋patients with colorectal cancer showed higher than HLA-A^＊0201＋healthy individuals [（ 2. 409 ± 2. 385） %,（ 0. 020 ±0. 021） % respectively],which was statistically significant（ P = 0. 008）. Conclusion： The frequencies of CAP-1-specific CD8＋T cells in PBMC from peripheral blood and lymph node of HLA-A^＊0201＋patients were increased,showed CEA-specific CTs has a vital role in colorectal cancer.