人补体杀伤小鼠胸腺瘤EL-4细胞的机制及其非致死模型的建立

Study on the mechanism of murine thymoma EL-4 cell induced human complement ac tivation and on the construction of it's sublytic model

目的探讨人血清 补体 杀伤小鼠胸腺瘤EL-4细胞的机制,建立补体杀伤T淋巴细胞的非致死模型。方 法用阻断补体活 化途径的方法探讨EL-4细胞活化人补体系统的机制,以MTT比色及LDH释放法绘制不同浓度 的人血清补体对EL-4细胞的裂解曲线,并以此确定非致死剂量。结果未致敏EL-4细胞可直接以Ca^(2+)、Mg^(2+)离子依赖的第一 途径活化人血清补体系统,在细胞数为1×10~4～2×10~4时,1∶200稀释的 人血清补体为EL-4细胞的非致死剂量。结论本研究 为探讨补体非致死性攻击在T淋巴细胞活化中的作用提供了良好的模型。多次实验结果表明 :本模型具有稳定、重复性好及剂量容易控制的优点,可供实验研究采用。

Objective To explore the mechanism of human serum complement in duced murine thymoma EL-4 cell lysis and to construct the complement nonleth al attack model on Tlymphocytes.Methods Complement activation was blocked to study the mechanisms of human serum complement mediated EL-4 cell lysis.The relationship between EL-4 cell lysis and complement dilution,and the sublyti c dose was determined with MTT colorimetric and LDH releasing assay.Results Human complement can be activated by unsensitized EL-4 cell through the first pa thway.When the cell number was 1×10~4～2×10~4,1∶200 diluted human serum was determined as the non-lethal dose. Conclusion Our study provided an excellent model for study the role o f non-lethal complement attack in Tlymphocytes activation,and it is confirm ed that the model are practical and stable,so can be used widely.