DC瘤苗激活的T淋巴细胞对胃癌细胞的杀伤效应研究

The killing effect of dendritic cell-tumor fusion vaccine stimulated T cells on SGC7901 gastric carcinoma cells

目的评估受DC融合性细胞瘤苗刺激的自体/异体T淋巴细胞的增殖性及其对肿瘤细胞的杀伤效应。方法收集健康志愿者外周血中白细胞层,经分离培养后获得成熟DC,再与SGC7901胃癌细胞经聚乙二醇(PEG)诱导融合,对融合细胞HAT/HT筛选培养,获得纯净的DC融合性细胞瘤苗。应用混合淋巴反应对DC融合性细胞瘤苗激活自体异体T淋巴细胞的增殖能力进行检测。选择DC融合性细胞瘤苗激活T淋巴细胞最强的1组,与SGC7901胃癌细胞混合培养,检测T淋巴细胞对肿瘤细胞的杀伤效性。结果200ml外周血制成的1U白膜可以获得(3.87±0.31)×107个DC细胞,成熟DC与SGC7901胃癌细胞经PEG诱导融合,可获得纯净DC融合性细胞瘤苗。DC融合性细胞瘤苗可显著刺激T淋巴细胞增殖,与自体/异体T淋巴细胞都是在效靶比10∶1时刺激能力最强,2种T淋巴细胞经DC融合性细胞瘤苗刺激后其增殖效应差异无统计学意义。DC融合性细胞瘤苗激活的自体/异体T淋巴细胞对SGC7901胃癌细胞的杀伤率分别为(77.16±2.76)%和(77.70±3.55)%,2种静息T淋巴细胞的杀伤率分别为(20.45±2.12)%和(21.31±2.86)%,DC融合性细胞瘤苗体外激活T细胞对SGC7901胃癌细胞的杀伤活性显著高于相应对照组(P<0.01)。结论采用健康志愿者DC制备的DC融合性细胞瘤苗在体外表现出对自体/异体T淋巴细胞具有相同的刺激效应,激活后的T淋巴细胞对肿瘤细胞的杀伤效率相似。

Objective To study the proliferation and the tumor cell killing of autologous and allogeneic T cells stimulated by dendritic cell-tumor fusion vaccine.Methods Mature dendritic cells were obtained from peripheral blood of healthy blood donors and fused with SGC7901 gastric carcinoma cells by polyethylene glycol,and then the dendritic cell-tumor fusion vaccine were screened out by HAT/HT medium.The proliferative activity of homogeneic and allogeneic T cells stimulated by dendritic cell-tumor fusion vaccine were detected.T cells with the strongest proliferative activity were isolated and co-cultured with SGC7901 gastric carcinoma cells,finally,the killing effect of T cells on tumor cells were detected.Results About（3.87±0.31）×10^7 dendritic cells were isolated from 200 ml peripheral blood,and purified dendritic cell-tumor fusion vaccine was obtained by fusing mature dendritic cells with SGC7901 gastric carcinoma cells.The dendritic cell-tumor fusion vaccine stimulated T cell proliferation effectively,and the proliferative activity of autologous and allogeneic T cells were the strongest at fusion cells to T cells ratio of 1∶10,meanwhile the proliferation activity of these T cells were similar.The killing rates of autologous and allogeneic T cells stimulated by dendritic cell-tumor fusion vaccine on SGC7901 gastric carcinoma cells were（77.16±2.76）% and（77.70±3.55）%,respectively.In contrast,the killing rates of autologous and allogeneic T cells were（20.45±2.12）% and（21.31±2.86）%,respectively,significantly lower than the vaccine activated T cells（P〈0.01）.Conclusion Dendritic cell-tumor fusion vaccine derived from dendritic cells of healthy blood donors can activate autologous and allogeneic T and effectively kill tumor cells in vitro.