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目的评价粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因修饰肿瘤疫苗的抗肿瘤活性,探讨其与化疗联合应用的抗肿瘤效果。方法以小鼠肺癌Lewis细胞系接种C57BL/6小鼠建立动物模型。利用含GM-CSF基因的重组质粒GM-CSF-pIRES2-EGFP,转染小鼠肺癌细胞系Lewis和LA795细胞,分别制备自体(Lewis细胞)和同种异体(LA795细胞)肿瘤疫苗。小鼠皮下接种Lewis细胞(1×107个)后5 d,采用GM-CSF分泌性肿瘤疫苗接种3次,同时设PBS组和单纯疫苗组作为对照,检测疫苗治疗前后脾细胞对Lewis细胞杀伤活性的变化,以及血清白介素4(IL-4)和γ干扰素(IFN-γ)的水平。观察GM-CSF分泌性肿瘤疫苗接种及联合化疗对小鼠生存期的影响。结果GM- CSF分泌性自体或同种异体肿瘤疫苗,均可诱导小鼠脾细胞对Lewis细胞杀伤活性增高,第3次接种后分别为(42.0±2.5)%和(39.6±7.3)%;同时血清中Th1类细胞因子IFN-γ的水平升高,而Th2类因子IL-4的水平无显著变化。GM-CSF分泌性肿瘤疫苗治疗组小鼠生存期与对照组比较,差异无统计学意义(P>0.05);而化疗联合疫苗组小鼠生存期较单独治疗组及对照组明显延长(P<0.05)。结论GM-CSF基因修饰的肿瘤疫苗可刺激机体产生特异性免疫反应;化疗的联合应用有助于提高肿瘤疫苗的治疗效果。
Objective To evaluate the antitumor activity of GM-CSF gene modified tumor vaccine and to evaluate the anti-tumor effect of GM-CSF gene modified tumor vaccine in combination with chemotherapy. Methods C57BL / 6 mice were inoculated with mouse lung cancer Lewis cell line to establish animal model. The lung cancer cell lines Lewis and LA795 were transfected with the recombinant plasmid GM-CSF-pIRES2-EGFP containing GM-CSF gene to prepare tumor vaccines of autologous (Lewis cells) and allogeneic (LA795 cells). The mice were subcutaneously inoculated with Lewis cells (1 × 107 cells) for 5 days and were inoculated three times with the GM-CSF secreting tumor vaccine. Meanwhile, the PBS group and the simple vaccine group were used as a control to detect the killing activity of the spleen cells against Lewis cells before and after the vaccine treatment , As well as the levels of serum interleukin 4 (IL-4) and interferon gamma (IFN-γ). To observe the effects of GM-CSF secreting tumor vaccination and chemotherapy on the survival of mice. RESULTS: GM-CSF secreting autologous or allogeneic tumor vaccine could induce the increase of cytotoxicity of Lewis cells in mice spleen cells after (42.0 ± 2.5)% and (39.6 ± 7.3)%, respectively Serum Th1 cytokine IFN-γ levels increased, while the level of Th2-like factors IL-4 no significant change. The survival time of mice in GM-CSF secreting tumor vaccine group was not significantly different from that in control group (P> 0.05). The survival time of mice in combination vaccine group was significantly longer than that in the control group and the untreated group (P < 0.05). Conclusion GM-CSF gene modified tumor vaccine can stimulate the body to produce specific immune response; the combination of chemotherapy can help to improve the therapeutic effect of tumor vaccine.