三阴乳腺癌细胞-树突状细胞融合疫苗的抗肿瘤免疫效应

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;ELISA试剂盒检测IL-12、IFN-γ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。

Antitumor effect of a triple negative breast cancer-dendritic cell fusion vaccine

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(IL-12) and interferon-γ(IFN-γ) 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 cytotoxicity of autologous T lymphocytes.Results The DCs isolated from peripheral blood mononuclear cells highly expressed CD83,CD86,CD11c 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 vaccine prepared by electrofusion can stimulate the proliferation of T lymphocytes and induces strong cytotoxicity of the T cells against breast cancer cells.