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目的 探讨IFN γ基因修饰胶质瘤瘤苗的可行性。方法 以重组腺病毒将鼠IFN γ(mIFN γ)基因转染小鼠G4 2 2胶质母细胞瘤细胞 (G4 2 2 /mIFN γ细胞 ) ,ELISA法检测该细胞中目的基因的表达 ,MTT法检测体外增殖能力 ,动物实验观察其体内成瘤性及抗肿瘤免疫功能变化 (实验组 )。设转染LacZ基因的病毒对照 (病毒对照组 )和野生G4 2 2细胞对照 (细胞对照组 )。结果 G4 2 2 /mIFN γ细胞能分泌高水平的mIFN γ (80 0 0pg/ml) ,和对照组相比 ,体外增殖能力无显著的差异 (P >0 0 5 ) ,皮下接种后小鼠存活期延长 ,有 6 6 7%的小鼠长期无瘤生存 (P <0 0 1)。免疫功能检测显示 ,实验组小鼠腹腔MΦ受LPS刺激后mTNFα (5 6 5 8± 2 6 4 6pg/ml)和NO (15 8 3± 2 1 6 μmol/L)分泌增高 ;脾NK、CTL及腹腔MΦ杀伤活性 (5 3 7± 7 4 %、 6 5 4± 7 4 %和 4 7 5± 5 9)显著增强 (P <0 0 1)。结论 mIFN γ基因修饰的G4 2 2细胞体内成瘤性降低 ,能诱导机体产生抗肿瘤免疫反应。
Objective To investigate the feasibility of IFN γ gene modified glioma vaccine. Methods MIFNγ gene was transfected into mouse G4 2 2 glioblastoma cells (G4 2 2 / mIFN γ cells) with recombinant adenovirus. The expression of the target gene was detected by ELISA. MTT assay The proliferation ability in vitro was tested. The animal experiment was conducted to observe the tumorigenicity and anti-tumor immune function (experimental group). The viral control (virus control) transfected with LacZ gene and the wild G422 cell control (cell control) were established. Results G4 2 2 / mIFN γ cells secreted high levels of mIFN γ (80 000 pg / ml), showing no significant difference in proliferation in vitro compared with the control group (P 0 05). Mice survived after subcutaneous inoculation Prolonged, 6 6 7% of mice long-term tumor-free survival (P lt; 0 0 1). Immunofluorescence showed that the MΦ in the peritoneal cavity of experimental mice was significantly increased after stimulation with LPS (56 65 ± 6 6 6 pg / ml) and NO (15 8 3 ± 2 1 6 μmol / L) And the peritoneal MΦ cytotoxicity (53 7 ± 74%, 654 ± 74% and 475 ± 59) were significantly increased (P <0.01). Conclusion The mIFN γ gene-modified G4 2 2 cells have reduced in vivo tumorigenicity and can induce the body to produce anti-tumor immune responses.