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作 者:张鹏[1] 姜华[1] 刘瑞磊[1] 刘宇[1] 张翘楚[1] 黄勇[1]
机构地区:[1]中山大学附属第三医院甲状腺乳腺外科,510630
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)
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.
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