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Ⅱ型单纯疱疹病毒(Herpes simplex virus 2,HSV-2)是引起生殖器疱疹(Genital herpes,GH)的主要病原体,研制有效的HSV-2疫苗是控制病毒传播的有效途径,本研究通过穿梭载体pDC316将糖蛋白D2(Glucoprotein D2,gD2)基因连接到腺病毒载体上,制备HSV-2重组腺病毒rAd-gD2,经PCR和Western blot的方法在基因和蛋白水平上鉴定正确后,分别于第0、2、4周免疫小鼠,以PBS和空白腺病毒载体(AdV)为对照,第7周取血,通过检测小鼠血清中gD2特应性IgG和中和抗体评价体液免疫应答,检测Th1/Th2型细胞因子评价细胞免疫应答。结果显示经过3次免疫,小鼠产生了高水平的gD2特应性IgG和较高水平的中和抗体,Th1型细胞因子IFN-γ和IL-2明显高于对照组,Th2型细胞因子IL-4、IL-5和IL-10均高于对照组,P<0.01,具有显著的统计学意义。本研究制备的rAd-gD2既可以刺激小鼠产生体液免疫应答,也可以产生细胞免疫应答,与预期设想一致,为未来HSV-2疫苗的研发提供了数据基础。
The herpes simplex virus 2 (HSV-2) is the major causative agent of genital herpes (GH). To develop an effective HSV-2 vaccine is an effective way to control the spread of the virus. In this study, the shuttle vector pDC316 The recombinant adenovirus vector was constructed by ligating the gene encoding Glucoprotein D2 (gD2) to the adenovirus vector. The recombinant adenovirus rAd-gD2 was prepared by PCR and Western blot. , And the mice were immunized with 2,4 and 4 weeks. PBS and blank adenoviral vector (AdV) were used as control. Blood was collected at the 7th week. The humoral immune response was evaluated by detecting gD2 atopic serum IgG and neutralizing antibody in mice serum. / Th2 cytokines to assess cellular immune responses. The results showed that after three immunizations, mice produced high levels of gD2 atopic IgG and higher levels of neutralizing antibodies, Th1-type cytokines IFN-γ and IL-2 were significantly higher than the control group, Th2 type cytokines IL -4, IL-5 and IL-10 were higher than the control group, P <0.01, with significant statistical significance. The rAd-gD2 prepared in this study can both stimulate the humoral and cellular immune responses in mice, which is in line with the expectation and provide a data basis for the future development of HSV-2 vaccine.