L1210细胞表面IL-2受体的表达和以IL-2为导向的免疫毒素活性的检测  

Expression of IL-2 Receptors on the Surfaces of L1210 Cells and Bioactivity Assay for IL-2 toxin Fusion Proteins

作　　者：张丽丽[1] 聂凌虎[1] 李焕娄[1] 李嗣英[1] 卢圣栋[1] 

机构地区：[1]中国医学科学院中国协和医科大学医药生物技术研究所药理室,北京100050

出　　处：《中国医学科学院学报》2000年第4期344-347,共4页Acta Academiae Medicinae Sinicae

摘　　要：目的　证明 L12 10细胞表面表达 IL- 2受体 (IL- 2 R) ,并以此为基础建立检测以 IL- 2为导向的免疫毒素的杀伤细胞活性的体内、体外实验模型。方法　采用免疫荧光染色及流式细胞测定技术 ,测定 L12 10细胞表面IL- 2 R。用 MTT法测定 IL- 2与两种由突变改型的绿脓杆菌毒素片段构建的两种融合蛋白 (IL- 2 - PEZNM和 IL- 2 -K- PEZNM)的体外杀伤细胞活性 ;以腹腔注射 L12 10细胞诱发小鼠腹水瘤 ,观察腹腔注射 IL- 2 - K- PEZNM对腹水瘤的抑制作用。结果　 86 .3%的 L12 10细胞表面表达 IL- 2受体 ,其荧光强度为 5 1.5 5 ;IL- 2 - PEZNM和 IL- 2 - K-PEZNM在体外对 L12 10细胞具有杀伤作用 ,且呈剂量效应相关 ,其半数抑制浓度 (IC5 0 )分别为 0 .5 1和0 .6 7μg/ ml;IL- 2 - K- PEZNM对小鼠腹水瘤生长具有较好的抑制作用 ,每只小鼠腹腔注射 5 μg/ d IL- 2 - K- PEZNM组生存时间较对照组延长 78.6 %。结论　 L12 10细胞是检测以 IL- 2为导向的免疫毒素体内外杀伤细胞活性的一种简便、稳定和安全的实验模型。Objective To confirm that L1210 cells surfaces express IL 2 receptors (IL 2R) and to establish experimental models for studying the cytotoxic effect of IL 2 toxin fusion protein in vivo and in vitro. Methods IL 2R on mouse leukemia L1210 cells was analyzed with immunofluorescence stain and FACS. The cytotoxic effects in vitro of these fusion proteins IL 2 PEZNM and IL 2 K PEZNM were assayed by MTT method; mice ascitic L1210 leukemia was induced by i.p. L1210 cells and the therapeutic effect of IL 2 K PEZNM was observed. Results 86 3% of L1210 cells expressed IL 2R on their surfaces and the relative fluorescence intensity was 51.55. Two fusion proteins IL 2 PEZNM and IL 2 K PEZNM showed dose related cytotoxicities to L1210 cells in vitro with the IC50 of 0.51 and 0.67 μg/ml respectively. IL 2 K PEZNM (5μg/d) showed siginificantly inhibitory effect on L1210 cells and prolonged the survival time of mice by 78 6%. Conclusions L1210 cells as a model system to measure the cytotoxic activity of IL 2 toxin fusion proteins both in vitro and in vivo is simpler,stabler, and safer model than those with other cells such as Hut102 and activated T lymphocytes.

关 键 词：L1210细胞 IL-2受体 融合蛋白 免疫毒素活性 

分 类 号：R392.12[医药卫生—免疫学]