IL-12协同B7-1诱导机体抗肿瘤免疫的实验研究

目的研究白细胞介素-12(Interleukin-12,IL-12)协同共刺激分子B7-1,联合诱导机体抗肿瘤免疫对大鼠卵巢上皮癌的治疗作用并探讨其机理.方法用携带有小鼠IL-12和B7-1的逆转录病毒表达载体感染大鼠卵巢癌细胞株NuTu-19,建立高表达细胞株NuTu-19/IL-12、NuTu-19/B7-1及双基因共表达细胞株NuTu-19/IL-12-B7-1,并以转染空载体pLXSN的细胞NuTu-19/Neo为对照.用经丝裂霉素C处理的各种基因修饰的肿瘤细胞免疫动物,观察卵巢癌腹腔转移模型动物生存期,及其诱导细胞毒性T淋巴细胞(cytotoxic lymphocyte,CTL)杀伤活性的作用.结果经各种基因修饰的肿瘤细胞免疫后,大鼠脾淋巴细胞增殖能力有不同程度的提高,CTL杀伤同源肿瘤细胞的活性明显增强,IL-12和B7-1基因联合修饰的肿瘤细胞免疫动物对模型动物生存期的延长具有显著意义(P＜0.05).结论IL-12和B7-1基因联合修饰的肿瘤细胞可以刺激机体CTL增殖、成熟,增强CTL对肿瘤细胞的识别和杀伤活性等,两者联合具有明显的协同效应.联合免疫基因治疗作为一种新的卵巢癌治疗方法,值得深入研究.

Study of the induction of anti-cancer immunity effects of interleukin 12 and co-stimulatory molecules B7-1

Objective To study the therapeutic effect and the synergetic mechanism of interleukin 12 (IL-12) and co-stimulatory molecule B7-1(CD80) on rat ovarian cancer model. Methods Retroviral vector pLmIL-12SN or pLmB7-1SN,which encoded mouse IL-12 or B7-1,was used to infect NuTu-19,rat epithelial ovarian cancer cell line,by lipofectin-mediated gene transfer system. Cell lines which could highly express either or both of IL-12 or B7-1,were named NuTu-19/IL-12,NuTu-19/B7-1 or NuTu-19/IL-12-B7-1 accordingly,and NuTu-19/Neo (transfected by vector pLXSN) was used as a control. The intraperitoneal disseminated ovarian cancer animal models were immunized with various mitomycin C-treated gene modified tumor cells. The survival time was observed,and the anti-tumor mechanisms were discussed. Results The splenic lymphocytes proliferation and cytotoxic effect of cytotoxic lymphocytes (CTLs) on syngeneic tumor cells increased significantly in fischer 344 rats immunized by various MMC -treated cytokine modified tumor cells,especially in IL-12 and B7-1 coexpressing group,which also had a prolonged survival time( P <0.05). Conclusion IL-12 and B7-1 gene modified tumor cells can induce anti-tumor immunity through stimulating lymphocytes proliferation and inducing recognition and cytotoxic effects of CTLs on tumor cells. An obvious synergistic effect exists when the two cytokines are combined,and it seems to be a promising therapeutic strategy for ovarian cancer.