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制各大鼠BERH-2肝癌细胞和激活B细胞融合肿瘤疫苗并观察其免疫相关表型,为融合肿瘤疫苗应用人体提供依据。方法:应用PEG在不同条件下,将肝癌细胞与激活B细胞融合。流式细胞仪鉴定融合细胞比例和检测MHC-/Ⅱ、B7、ICAM-1、LFA-1的水平。结果:50%PEG-1400介导细胞融合60秒获得融合细胞百分比最高。与融合前BERH-2细胞相比,融合细胞获得了MHC-Ⅱ、B7、ICAM-1、LFA-1的表达,MHC-I表达增加。结论:制备大鼠BERH-2肝癌细胞和激活B细胞融合肿瘤疫苗的最佳条件为50%PEG-1400融合60秒。融合细胞不但表达肿瘤抗原,同时表达免疫原性相关分子。
Preparation of rat BERH-2 hepatocellular carcinoma cells and activated B cell fusion tumor vaccine and observe its immune-related phenotypes, provide a basis for the integration of tumor vaccine application of human body. Methods: PEG was used to fuse hepatoma cells with activated B cells under different conditions. Flow cytometry was used to identify the proportion of fused cells and to detect the levels of MHC-/II, B7, ICAM-1, and LFA-1. Results: The percentage of fused cells was highest when 50% PEG-1400 mediated cell fusion for 60 seconds. Compared with pre-fusion BERH-2 cells, fused cells obtained expression of MHC-II, B7, ICAM-1, LFA-1 and increased expression of MHC-I. CONCLUSION: The best condition for preparing rat BERH-2 hepatoma cells and activating B cell fusion tumor vaccine is 50% PEG-1400 fusion for 60 seconds. Fused cells express not only tumor antigens but also immunogenic related molecules.