癌-睾丸抗原OY-TES-1致敏的树突状细胞体外抗胶质瘤的免疫效应

Immune Effect against Glioma Mediated by Dendritic Cells Pulsed with Cancer-testis Antigen OY-TES-1 In Vitro

目的研究负载癌-睾丸抗原(cancer-testis antigen,CTA)OY-TES-1的树突状细胞(dendritic cell,DC)体外抗胶质瘤的细胞免疫效应。方法细胞因子联合诱导HLA-A2+健康人外周血单核细胞为DC,采用OY-TES-1重组蛋白致敏DC,流式细胞术检测成熟DC表型分子;进而使DC诱导同体T淋巴细胞产生细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL),CCK8检测T细胞增殖情况,ELISPOT检测IFN-γ含量,乳酸脱氢酶释放法检测CTL对胶质瘤细胞U251和A172的杀伤作用。结果负载OY-TES-1的DC低表达CD14,而高表达CD1a、CD83和CD80表型分子,能明显促进CD8+T细胞的增殖及其IFN-γ的分泌(P<0.05);而OY-TES-1特异性CTL在效靶比为50∶1时对HLA-A2+和OY-TES-1+U251、HLA-A2-和OY-TES-1+A172均有明显杀伤作用(均P<0.05),但其对U251的杀伤作用更强。结论体外用OY-TES-1重组蛋白致敏的DC可诱导特异性抗胶质瘤免疫应答,提示OY-TES-1可能成为胶质瘤免疫治疗的靶点。

Objective To detect immune effect against glioma mediated by dendritic cells(DCs)pulsed with cancer-testis antigen(CTA)OY-TES-1 in vitro.Methods Cytokines induced HLA-A2+healthy human peripheral blood mononuclear cells into DCs,which were loaded by OY-TES-1 recombinant protein.Then flow cytometry was used to detect the phenotype molecules of mature DC.T cell proliferation was tested by CCK8,and IFN-γlevel was assessed by ELISPOT assay.Lactate dehydrogenase release method was used for the killing effect of CTL on glioma cells U251 and A172.Results DCs loaded OY-TES-1 showed the low expression of CD14 and high expression of CD1 a,CD83 and CD80,and could significantly promote the proliferation of CD8+T cells and the secretion of IFN-γ(P<0.05).OY-TES-1 specific CTL had significant killing effect on HLA-A2+and OY-TES-1+U251 and HLA-A2-and OY-TES-1+A172 at the target-target ratio of 50∶1(P<0.05),but its killing effect was much stronger on U251 than on A172.Conclusion DCs loaded OY-TES-1 can induce specific anti-glioma immune response,suggesting that OY-TES-1 may be the target for glioma immunotherapy.