重组人IL-18在大肠杆菌中的表达及其抗肿瘤作用  

The expression and anti-tumor activity of rhIL-18

作　　者：张海峰[1] 盛伟华[1] 缪竞诚[1] 谢宇锋[1] 李丽娥[1] 马丽丽[1] 庄文卓[1] 杨吉成[1] 

机构地区：[1]苏州大学医学院细胞和分子生物学教研室,苏州215007

出　　处：《中国免疫学杂志》2006年第2期141-144,共4页Chinese Journal of Immunology

基　　金：江苏省高校自然科学研究项目(01KJB180001)

摘　　要：目的:获取rhIL-18原核表达产物并研究其活性。方法:用本室构建的含基因重组表达载体PBV220-IL-18的大肠杆菌DH5α,经42℃热诱导表达和包涵体提取纯化后获取rhIL-18蛋白;采用ELISA试剂盒检测rhIL-18刺激PBMC分泌IFN-γ的活性及MTT法检测其对NK细胞杀伤K562细胞自然杀伤活性;同时用荷瘤小鼠模型检测其体内抗瘤功能。结果:诱导含重组表达载体PBV220-IL-18的大肠杆菌DH5α后,蛋白电泳显示出一条相对分子量(Mr)为18 000的蛋白条带;10μgrhIL-18蛋白体外刺激PBMC后,其分泌IFN-γ的能力与对照组相比提高了8倍,细胞毒性提高3倍;同时rhIL-18蛋白能明显抑制肿瘤生长和延长荷瘤鼠生存期。结论:获得了有免疫调节功能和抗肿瘤活性的rhIL-18蛋白,为今后IL-18重组产物的研制和开展肿瘤的生物治疗提供了实验依据。Objective：To obtain prokaryotic expression protein of IL-18 and research its function. Methods：By heat-inducing E. coli DH5α carrying expression vector PBV220-IL-18 which were constructed by us, and by extracting and purifing inclusion body, the protein of IL-18 was obtained,then the fimction of promoting IFN-γ secretion from PBMC is tested by ELISA ,the cytotoxity of the NK cells to K.562 cells was tested by MTI＇, and the anti-tumor function in vivo was tested by mice tumor model. Results：After inducing E. coli DH5 at carrying expression vector PBV220-IL- 18, the protein with relative molecular mass （Mτ） 18 000 was detected ; after stimulated by 10μg rhIL-18 protein in vitro, IFN-γ, secretion and the cytotoxity of PBMC comparing with the control group was promoted by 8 times and 3 times respectively,and the average survival time of mice carrying tumor was obviously prolonged. Conclusion：The rhIL-18 protein which had the immune regulation function and anti-rumor activity was obtained ,so that provide an experiment basis for futher research and manufacture and tumor biologic therapy of rhIL-18.

关 键 词：白介素18 基因表达 免疫调节功能 抗肿瘤活性 

分 类 号：R392.12[医药卫生—免疫学]