狂犬病毒糖蛋白DNA疫苗的构建及免疫效果的研究

Construction and Immune Effect of Rabies Glycoprotein DNA Vaccine

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 rabies glycoprotein DNA vaccine and study its immune effect in mice. Methods Isolate glycoprotein gene from rabies virus CTN strain, amplify by RT-PCR, identify by sequencing, then insert into VR1055 vector. Extract the recombinant plasmid in a large quantity by alkaline lysis, purify by Sepharose 4FF column chromatography and inject i.m. into NIH mice. Determine the humoral immunity of mice by ELLSA, and cellular immunity by MTT method. Results VR1055-CTN DNA vaccine induced antibody against rabies virus. Both the spleen lymphocyte transformation index and IL-2 activity in sera of mice immunized with VR1055-CTN DNA vaccine were significantly higher than those immunized with empty vector. After challenge with rabies virus CVS strain, the survival rates of mice immunized with VR1055-CTN DNA vaccine and empty vector were 73% and 25% respectively. Conclusion The VR1055-CTN DNA vaccine containing rabies virus glycoprotein gene induced both humoral and cellular immunity and provided good immune protection.