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目的:评价HA纳米载体转hGM-CSF基因的HepG2疫苗的抗肿瘤活性。方法:SCID小鼠20只腹腔内注射健康志愿者外周血淋巴细胞(1×107/ml)1.0 ml,同时背部皮下接种人肝癌HepG2细胞(1×107ml)0.2 ml。当皮下移植瘤体积长至100 mm3时,随机分四组:Ⅰ组,60Co照射的转染GM-CSF基因的HepG2细胞组,Ⅱ组,60Co照射的HepG2细胞组,Ⅲ组,生理盐水组,Ⅳ组,接种前,每组5只。MTT法检测各组小鼠脾细胞CTL活性,ELISA法检测血清IL-4和IL-12的水平。结果:转染GM-CSF基因的HepG2疫苗组小鼠脾细胞CTL活性明显高于其余组(P<0.05);同时血清Th1类细胞因子IL-12的水平明显升高(P<0.05),而Th2类细胞因子IL-4的水平无统计学意义(P>0.05)。结论:HA纳米载体转hGM-CSF基因的HepG2疫苗可刺激机体产生特异性免疫反应。
Objective: To evaluate the antitumor activity of HepG2 vaccine with hGM-CSF gene transferred from HA nanocarriers. METHODS: Twenty SCID mice were intraperitoneally injected with 1.0 ml of peripheral blood lymphocytes (1×107/ml) in healthy volunteers, and 0.2 ml of human hepatoma HepG2 cells (1×107 ml) were subcutaneously inoculated on the back. When the subcutaneous tumor size increased to 100 mm3, they were randomly divided into four groups: Group I, HepG2 cells transfected with GM-CSF gene irradiated with 60Co, Group II, HepG2 cells irradiated with 60Co, Group III, and saline groups. Group IV, before inoculation, 5 in each group. The activity of CTL in spleen cells of each group was detected by MTT assay, and the levels of serum IL-4 and IL-12 were detected by ELISA. RESULTS: The CTL activity of spleen cells in HepG2 vaccine group transfected with GM-CSF gene was significantly higher than that in the other groups (P<0.05), and the level of serum Th1 cytokine IL-12 was significantly increased (P<0.05). The level of Th2 cytokine IL-4 was not statistically significant (P>0.05). CONCLUSION: HepG2 vaccine with HA nanocarriers transfected with hGM-CSF gene can stimulate the body to produce specific immune responses.