【摘要】 目的:探讨HLA-A2-的黑色素瘤细胞与HLA-A2+的树突状细胞(DC)融合后体外诱导Me-lan-A特异性细胞毒性T淋巴细胞(CTL)的作用及交叉抗原递呈的能力。方法:用PEG法将黑色素瘤细胞与DC融合,在含GM-CSF的RPMI1640完全培养基条件下继续培养24～48h,然后将融合细胞与Me-lan-A特异性T细胞共同培养,用细胞内细胞素染色法检测其诱导CTL的活性,流式细胞术分析T细胞的活化率。结果:从异体外周血分离的单核细胞在GM-CSF及IL-4条件下培养6d后,未成熟的DC能表达一定的CD80,CD83,CD86,HLA-DR和HLA-ABC等共刺激分子,而经TNF-浕,PGE-2及CD40L诱导成熟的DC,则这些分子的表达进一步上调。融合细胞体外活化Melan-A特异性T细胞的效率为16.72%±4.26%,阴性对照为0.21%±1.84%,阳性对照为28.60%±5.67%。融合细胞活化的CTL能有效地溶解HLA-A2的黑色素瘤细胞。结论:HLA-A2-的黑色素瘤细胞与HLA-A2+阳性的DC融合肿瘤疫苗,能在体外有效地交叉递呈MHC-I限制性肿瘤抗原,并诱导Melan-A特异性的CTL和有效地杀伤黑色素肿瘤细胞。更多还原

【Abstract】 Objective To determine the effect of activation of specific anti-tumor cytotoxic T lymphocytes (CTL) and the ability of cross-presentation in vitro by fusion of HLA-A2+ human dendritic cells (DCs) with HLA-A2-melanoma cells. Methods The HLA-A2+ human dendritic cells and HLA-A2-melanoma cells were fused by PEG and were cultivated in complete RPMI1640 media containing FCS (10%) and GM-CSF for 24~48 h,and then co-cultured fusion cells with Melan-A specific T cells. HLA-A2-melanoma cells were negative control,While T2 cells and DC+Pts were positive control. The activation of anti-tumor CTL elicited by the fusion cells was detected by intracellular cytokine staining. Results The immature DC could express CD80,CD83,CD86,HLA-DR,and HLA-ABC,but the mature DC induced by TNF-α,PGE-2,and CD40L further highly expressed above molecules. The rate of specific CTL cells primed by the fusion cells was 16.72%±4.26%,negative control was 0.21%±1.84%,and positive control was 28.60%±5.67%. The CTL from vaccine by fusing DC and LAR6 induced lysis of HLA-A2+ LAR1 cells. Conclusion The HLA-A2 restricted specific anti-tumor CTL can be induced in vitro by fusion of HLA-A2+ human dendritic cells with HLA-A2-melanoma cells.更多还原