三阴乳腺癌细胞-树突状细胞融合疫苗的抗肿瘤免疫效应

摘要

目的 利用细胞电融合技术制备外周血来源树突状细胞(DC)和三阴乳腺癌细胞(MDA-MB-231)全抗原肿瘤疫苗,观察其特异性抗肿瘤免疫效应.方法 从健康人外周血中分离培养DC,利用电融合技术,将DC和三阴乳腺癌细胞融合；荧光显微镜观察融合疫苗的形态；流式细胞仪进行融合疫苗的表型鉴定；ELlSA试剂盒检测IL-12、IFN-y的分泌情况；CCK-8试剂盒测定融合疫苗刺激同源异体T淋巴细胞增殖和细胞毒性效应.结果 成功分离培养DC,其表面高表达DC的分子标记CD83、CD11c、CD86、HLA-DR;融合疫苗形态不规则,其表面共同表达DC和三阴乳腺癌的标记分子；T淋巴细胞增殖实验证明融合疫苗有很强的免疫刺激活性；细胞毒性实验证明融合疫苗具有比对照组更强的杀伤肿瘤细胞作用.结论 电融合技术成功诱导DC和三阴乳腺癌细胞融合,全抗原融合疫苗体外实验可显著增强抗肿瘤免疫效应.%Objective To test the antitumor effect of a human triple-negative breast cancer cell-dendritic cell (DC) fusion vaccine.Methods DCs were isolated from fresh peripheral blood of healthy donors.The fusion vaccine was prepared by fusing the DCs and MDA-MB-231 cells via electrofusion.The morphology of the vaccine was indentified under inverted fluorescence microscope and the phenotypes were analyzed with flow cytometry.The production of interleukin-12 (JL-12) and interferon-y (IFN-y) by the fusion cells was assessed using ELISA.A CCK-8 kit was used to examine the effect of the vaccine in stimulating the proliferation and cytotoxiciry of autologous T lymphocytes.Results The DCs isolated from peripheral blood monomiclear cells highly expressed CD83,CD86,CDllc and HLA-DR on the cell surface.The fusion cells were irregular in shape and coexpressed the phenotypes of DCs and MDA-MB-231 cells.The fusion cells possessed a strong ability to stimulate the proliferation of T lymphocytes in vitro.Compared with the control group,the fusion vaccine showed a stronger antitumor effect against the breast cancer cells.Conclusion The triple-negative breast cancer-DC fusion vacdne prepared by electrofusion can stimulate the proliferation of T lymphocytes and induces strong cytotoxidty of the T cells against breast cancer cells.