糖基化磷脂酰肌醇修饰的小鼠IL-21瘤苗抗肿瘤效应及其机制初探  

Investigation on the antl-tumor effect and mechanisms of the glycosyl phosphatidylinositol modified routine IL-21 tumor vaccine

作　　者：赵枫姝[1] 褚莉莉[1] 窦骏[1] 唐权[1] 王永仿[1] 吴昀[1] 曹明刚[1] 潘猛[1] 顾宁[2] 

机构地区：[1]东南大学基础医学院病原生物学和免疫学系,南京210009 [2]东南大学生物科学与医学工程学院

出　　处：《中华微生物学和免疫学杂志》2008年第9期822-827,共6页Chinese Journal of Microbiology and Immunology

基　　金：江苏省六大人才高峰项目（医药行业D14）;国家自然科学基金项目（90406023）

摘　　要：目的构建糖基化磷脂酰肌醇（glycosyl phosphatidylinositol，GPI）修饰的小鼠IL-21瘤苗，并对此瘤苗的抗肿瘤效应及其机制作初步探讨。方法通过重叠PCR方法获得IL-21-GPI融合基因并将其插入空载体pcDNA3．1。将鉴定过的重组载体以脂质体法转染B16F10细胞制成瘤苗，细胞间接免疫荧光法及流式细胞仪检测转染瘤细胞膜表面IL-21的表达，通过对小鼠脾细胞的增殖作用鉴定表达的IL-21的生物学活性。将瘤苗接种小鼠后，通过观察小鼠肿瘤体积和生存率分析瘤苗的抗瘤性，并检测了瘤苗免疫鼠的细胞免疫活性。结果正确构建了pcDNA3．1／IL-21-GPI重组载体，膜表达的IL-21有良好的生物学活性，制备的瘤苗能发挥抗肿瘤效应，其机制与免疫鼠细胞免疫活性增强有关。结论成功构建了具有抗肿瘤活性的GPI修饰的IL-21瘤苗，为其进一步抗肿瘤免疫治疗研究奠定了基础。Objective To construct the murine IL-21 （ mIL-21 ） tumor vaccine modified by glycosyl phosphatidylinositol （GPI）, and to evaluate its anti-tumor effect and mechanisms. Methods The IL-21- GPI gene was acquired by overlap PCR and inserted into pcDNA3.1. The recombinant plasmid pcDNA3.1/ IL-21-GPI was transformed into cell B16F10, and the expression of raiL-21 on cell membrane was determined by cell indirect immumofluorescence and flow cytometry （FCM）. The bioactivity of mIL-21 was identified according to its effects on the proliferation of mouse spleen cells. The anti-tumor effect was evaluated depending on the tumor size and the survival of tumor-bearing mice after the tumor vaccine was inoculated into C57BL/6 mice. And the activity of cell-mediated immunity in immunized mice was detected at the same time. Results The recombinant plasmid pcDNA3.1/IL-21-GPI was correctly constructed, which could express mIL-21 binding the membrane with good bioactivity. The vaccine had good anti-tumor effect, and the cell-mediated immunity had been improved in immunized mice. Conclusion The GPI modified mIL-21 tumor vaccine with anti-tumor activity was constructed successfully, which provided a good foundation for studying anti-tumor immunity and therapy in future.

关 键 词：小鼠IL-21 糖基化磷脂酰肌醇 抗肿瘤免疫 

分 类 号：R73-36[医药卫生—肿瘤]