摘要
目的：利用细胞融合技术制备外周血来源树突状细胞(Dendritic cells, DCs)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: 成功分离培养树突状细胞，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。
Abstract
Objective: To test the effect of a human triple-negative breast cancer cell-dendritic cell(DC) fusion 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 analyzed by flow cytometry (FCM). The effect of the vaccine in stimulating the proliferation of autologous T lymphocytes was assessed using CCK-8 kit. Results: The DCs isolated from peripheral blood mononuclear cells highly expressed CD86, CD11c, 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 proliferation of T lymphocytes in vitro. Conclusion: The triple-negative breast cancer-DC fusion vaccine prepared by electrofusion can strongly stimulate the proliferation of T lymphocytes in vitro.
张 鹏;姜 华;刘瑞磊;刘 宇;张翘楚;黄 勇.
三阴乳腺癌疫苗刺激T淋巴细胞增殖免疫效应观察[J]. 新医学, 2012, 43(12): 861
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