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目的:建立稳定表达小鼠白细胞介素21(mIL-21)的肿瘤细胞瘤苗,观察其在小鼠体内是否能够诱导有效的抗肿瘤免疫反应及免疫记忆效应。方法:将已鉴定的重组质粒pcDNA3.1/mIL-21用脂质体法转染Sp2/0细胞制备瘤苗,RT-PCR法鉴定瘤苗中mIL-21的表达。通过流式细胞仪检测细胞周期来反映瘤苗体外增殖活性,再将其接种BALB/c小鼠,监测肿瘤生长情况,观察mIL-21瘤苗诱导的抗肿瘤效应;用ELISA法检测血清IFN-γ和IL-4含量。结果:得到稳定表达mIL-21的瘤苗Sp2/0-mIL-21。与对照组相比,体外增殖活性无差异。皮下接种BALB/c小鼠后,肿瘤生长缓慢,部分小鼠无瘤体生长并长期存活;用野生株Sp2/0瘤细胞再次攻击未长肿瘤的实验小鼠,4周后亦未见肿瘤生长。接种瘤苗小鼠血清中IFN-γ水平明显上升,IL-4无明显增高。结论:成功构建了mIL-21瘤苗Sp2/0-mIL-21,其能诱导有效的抗肿瘤免疫反应及免疫记忆效应。
OBJECTIVE: To establish a tumor cell vaccine stably expressing mouse interleukin-21 (mIL-21) and observe whether it can induce effective anti-tumor immune response and immune memory effect in mice. Methods: Sp2 / 0 cells were transfected with the identified recombinant plasmid pcDNA3.1 / mIL-21 by Lipofectamine 2000. The expression of mIL-21 in the vaccine was identified by RT-PCR. The cell cycle was detected by flow cytometry to reflect the in vitro proliferation activity of the tumor vaccine. The BALB / c mice were inoculated with the tumor cell growth to monitor the tumor growth. The anti-tumor effect induced by the mIL-21 vaccine was observed. The serum IFN- γ and IL-4 content. Results: The tumor vaccine Sp2 / 0-mIL-21 stably expressing mIL-21 was obtained. Compared with the control group, in vitro proliferation activity no difference. After inoculation BALB / c mice subcutaneously, the tumors grew slowly, some of the mice did not grow in the tumor and survived for a long time. After the mice were re-challenged with wild type Sp2 / 0 tumor cells, the mice without tumor grew 4 weeks later. . The level of IFN-γ in sera of mice inoculated with tumor vaccine increased obviously, and the level of IL-4 was not increased obviously. Conclusion: The mIL-21 tumor vaccine Sp2 / 0-mIL-21 was constructed successfully, which can induce effective anti-tumor immune response and immune memory effect.