论文部分内容阅读
目的构建狂犬病毒糖蛋白基因DNA疫苗,并检测其免疫小鼠的免疫应答。方法用RT-PCR法扩增和分离CTN株狂犬病毒糖蛋白基因,测序后克隆入VR1055载体中,构建重组质粒VR1055-CTN,经碱裂解法提取质粒,并经Sepharose4FF柱层析纯化后免疫NIH小鼠,用ELISA法检测体液抗体,用MTT法检细胞免疫,并进行常规效力试验。结果VR1055-CTNDNA疫苗具有较好的诱导狂犬病毒抗体的能力,在诱导细胞免疫中,刺激脾淋巴细胞转化指数和小鼠血清中的IL-2活性均高于空白载体对照,差异有显著意义(P=0.000)。效力试验的小鼠存活率分别为73%和25%。结论该疫苗既能诱导体液免疫,又能诱导细胞免疫,并具有较好的免疫保护效果。
Objective To construct the DNA vaccine of rabies virus glycoprotein gene and test its immune response. Methods The rabies virus glycoprotein gene of CTN strain was amplified by RT-PCR and cloned into VR1055 vector. The recombinant plasmid VR1055-CTN was constructed. The plasmid was isolated by alkaline lysis and purified by Sepharose 4 FF column. Mice were tested for humoral antibody by ELISA, cell immunization by MTT method, and routine efficacy test. Results The VR1055-CTN DNA vaccine had better ability of inducing rabies virus antibody. The induction of spleen lymphocyte transformation index and the activity of IL-2 in the serum of mice were higher than that of blank vector control in inducing cellular immunity (the difference was significant P = 0.000). The efficacy test mice survival rates were 73% and 25%. Conclusion The vaccine can not only induce humoral immunity but also induce cellular immunity with good immune protection.