摘要
目的:利用细胞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,观察其刺激同源T淋巴细胞增殖作用。方法:从健康人的外周血中分离培养树突状细胞,利用电融合技术,将DC和三阴乳腺癌细胞融合;用荧光显微镜观察融合疫苗的形态;用流式细胞仪进行融合疫苗的表型鉴定;用CCK-8试剂盒测定融合疫苗刺激同源异体T淋巴细胞增殖效应。结果:成功分离培养DC,其表面高表达DC的分子标记CD11c、CD86、HLA-DR;融合疫苗的形态不规则,其表面共同表达DC和三阴乳腺癌的标记分子;T淋巴增殖实验证明融合疫苗具有很强的免疫刺激活性。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同源T淋巴细胞增殖。
Objective： To test the effect of a human triple-negative breast cancer cell-dendritic cell （DC） fu- sion vaccine on the proliferation of T lymphocytes in vitro. Methods： DCs were isolated from fresh peripheral blood of healthy donors. The fusion cell vaccine was prepared by fusing the DCs and MDA-MB-231 cells by electrofusion. The morphology of the vaccine was indentified under inverted fluorescence microscope and the phenotypes were ana- lyzed by flow cytometry （FCM）. The effect of the vaccine in stimulating the proliferation of autologous T lympho- cytes was assessed using CCK-8 kit. Results： The DCs isolated from peripheral blood mononuclear cells highly ex- pressed CD86, CD1 lc, and HLA-DR on the cell surface. The fusion cells were irregular in shape and coexpressed the phenotypes of both DCs and MDA-MB-231 cells. The fusion cells showed a strong ability to stimulate the prolif- eration of T lymphocytes in vitro. Conclusion： The triple-negative breast cancer-DC fusion vaccine prepared byelectrofusion can strongly stimulate the proliferation of T lymphocytes in vitro.
出处
《新医学》
2012年第12期861-865,F0003,共6页
Journal of New Medicine
基金
中央高校基本科研业务费专项基金(09ykpy15)
关键词
三阴乳腺癌
树突状细胞
电融合
融合疫苗
Triple negative breast cancer
Dendritic cell
Electrofusion
Fusion vaccine
