Ad5介导mGM-CSF基因修饰A549细胞肿瘤疫苗的实验研究

Experimental Study on Ad5-Mediated mGM-CSF Gene Modified A549 Tumor Cell Vacccines

【目的】探讨Ad5介导mGM-CSF基因修饰A549细胞肿瘤疫苗在模型小鼠的有效性。【方法】Ad5/mGM-CSF感染体外培养的A549细胞(MOI=20),24 h后将A549细按照7×106细胞/0.1 mL/支分装冻存,20000rad照射灭活,溶化后直接用于注射;4周龄雄性C57小鼠,左腋下皮下接种A549细胞3 d后开始在右腋下皮下接种主动免疫制剂1支,以单纯经过照射的同等剂量的A549细胞为对照,每周1次,连续5周,每3 d记录肿瘤发生率、测量瘤体积。【结果】Ad5/mGM-CSF能有效的感染A549细胞,其表达量为524ng/106cell/24 h;肿瘤细胞+GM-CSF主动免疫疗法能够有效抑制皮下接种肿瘤的发生和增长。【结论】Ad5介导mGM-CSF基因修饰A549细胞肿瘤疫苗能有效的抑制A549细胞的增殖,为临床应用打下一定的基础。

[Objective] To explore the availability of Ad5-mediated mGM-CSF gene modified A549 tumor cell vaccines on mouse model. [Methods] Ad5/mGM-CSF was infected into A549 cells in vitro（MOI= 20）. Infected A549 cells were subpackaged and cryopreserved according to 7 × 10^6 cells/0, lmL/ramus after 24 hours later, and inactivated with 20000rad irradiating, and used directly for injection after colliquated. 4-week-old male C57 mouse was inoculated A549 cells subcutaneouly at left armpit, and inoculated one active immunity preparation subcutaneouly at right armpit after 3 days later, as compared with A549 cells exposed to isodose per week and continuously for 5 weeks. Tumorigenesis rate and tumour volume were noted. [Results] Ad5/ mGM-CSF was effectively infected into A549 cells with expression by 524ng/10^6cell/24 hours. Tumor cells plus GM-CSF active immunotherapy effectively suppressed the generation and growth of hypodermically inoculated tumor. [Conclusion] Ad5-mediated mGM-CSF gene modified A549 tumor cell vaccines could effectively suppress proliferation of A549 cells.