冻融肿瘤细胞和肿瘤细胞培养上清对淋巴细胞活化能力的影响

 目的　比较冻融肿瘤细胞和肿瘤细胞培养上清对淋巴细胞活化能力影响的差异。方法　常规冻融法制备恶性黑色素瘤B16细胞全肿瘤细胞抗原，收集培养不同时间的肿瘤细胞培养上清；利用Transwell法检测冻融肿瘤细胞以及肿瘤细胞培养上清对淋巴细胞的趋化能力；用CCK-8法检测各组趋化淋巴细胞的肿瘤杀伤活性。结果　冻融肿瘤细胞及培养2 h以上的肿瘤上清液对淋巴细胞均有趋化作用；培养4 h以上的肿瘤上清液的趋化能力明显强于冻融瘤细胞。各组趋化的淋巴细胞均具有肿瘤杀伤活性；培养4 h以上的肿瘤上清液组淋巴细胞的杀伤能力明显强于冻融肿瘤细胞组。在一定范围内，培养上清对淋巴细胞的趋化和活化能力随时间延长而增强。结论　培养一定时间的肿瘤上清液对淋巴细胞的趋化和活化能力均强于冻融肿瘤细胞，用培养上清液代替冻融肿瘤细胞抗原作为抗原活化淋巴细胞可获得更好的免疫效果。

Effect of the activation of lymphocyte between freeze-thaw tumor cell and the supernatant of tumorcell culture

Objective To compare the effect of freeze-thaw tumor cells and the supematant from tumor cell culture on the activation of lymphocytes. Methods Malignant melanoma B16 cells were prepared as tumor cell vaccine and the supernatant from tumor cell culture was collected at different time point. Transwell method was used to determine the ehemotaxis of lymphocyte attracted by freeze-thaw tumor cells and the supernatant from tumor cell culture. The cytotoxie activity of lymphocyte was detected by CCK-8 method. Results Freeze-thaw tumor cells and the supernatant from more than 2 h of tumor cell culture were found to show chemotaxis of lymphocyte. The ehemotaxis of tumor cell culture more than 4 h was stronger than freeze-thaw tumor cells. Each group of chemotactic lymphocytes demonstrated to have the activity of killing tumor cells. The ability of killing tumor cells induced by the tumor cell culture more than 4 h was stronger than that induced by freeze-thaw tumor cells. In a certain range, the ability of lymphocyte chemotaxis and activation were enhanced over time. Conclusion The chemotaxis and eytotoxic activation of lymphocyte induced by the supernatant from tumor cell culture for a certain time are stronger than those by freeze-thaw tumor cells. The supernatant from tumor cell culture can be used as tumor antigen to get better immune activation instead of the freeze-thaw tumor cells.