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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 investigate the mechanism of human serum complement in killing mouse thymoma EL-4 cells and to establish a non-lethal model of complement-killing T lymphocytes. Methods The mechanism of activating complement of human EL-4 cells was explored by blocking the pathway of complement activation. The cleavage curves of EL-4 cells by different concentrations of human serum complement were drawn by MTT colorimetry and LDH release method, Lethal dose. Results The non-sensitized EL-4 cells could directly activate the human complement complement system by Ca 2+ and Mg 2+ ion-dependent first pathway. The number of cells was 1 × 10 4 ~ 2 × 10 ~ At 4, human serum complement at a dilution of 1: 200 is a non-lethal dose of EL-4 cells. Conclusions This study provides a good model for investigating the role of complement non-lethal challenge in T lymphocyte activation. Multiple experimental results show that: The model has the advantages of stable, good repeatability and easy dosage control for experimental study.