编码糖蛋白B的裸基因疫苗诱导单纯疱疹病毒Ⅰ型感染小鼠的体液和细胞免疫

Naked gene vaccine of encoding glycoprotein B induces humoral and cellular immunity of herpes simplex virus-Ⅰ infection in mice

[目的]用编码单纯疱疹病毒Ⅰ型(HSV-1)糖蛋白B的pDNA转染小鼠,研究基因疫苗治疗感染性疾病的有效性,并分析免疫反应.[方法]采用血管内注射方法,将含有不同剂量、不同pDNAs的生理盐水注入Balb/c小鼠体内,1周后行再次免疫,观察以不同组合pDNA进行免疫的实验组和对照组小鼠被致死剂量HSV-1感染后生存率间的差异.通过潜伏感染的检测、血清中细胞因子水平的测定、细胞毒性反应的测定及中和实验评价免疫效果.[结果]接种pG.gB或接种pGEG.gB的小鼠,与相应对照组比较,生存率和诱导产生中和抗体的效价与剂量呈正相关,pGEG.gB免疫组对P815.gB细胞有显著的细胞毒性作用,pGEG.gB免疫小鼠的脾细胞增殖反应显著强于pG.gB免疫的小鼠;pG.gB和pGEG.gB免疫后的小鼠血清中IL-12和IFN-γ水平均显著升高.[结论]裸pDNA的经小鼠尾静脉免疫可诱导HSV-1感染小鼠的体液和细胞免疫应答,对小鼠HSV-1急性感染有明显的预防和治疗作用,且含有EBNA1和oriP的EBV质粒载体明显增强CTL活性和增殖反应.

OBJECTIVE The mice were transfected by injecting pDNA encoded as herpes simplex virus-Ⅰ(HSV-1) gB through tail vein to testify efficiency of treating infectious diseases with gene vaccine and analyze antiviral immune response induced by gene vaccine.METHODS The Balb/c mice were injected by normal saline with different dosages and pDNAs and immune again a week later,and the different survival rate was observeed in experimental and control group animals implemented immunization by different combination of pDNAs of infecting fatal dosage of HSV-1,and immunological effects were evaluated by detecting latent infection,cytokine concentrations of serum,cytotoxic reaction and neutralization test.RESULTS The mice inoculated with pG.gB or pGEG.gB were compared with the corresponding control group,the survival rate and induced neutralizing antibody titers were positive correlation with the measurement,pGEG.gB immune group had obvious cytotoxicity to P815.gB cells comparing to the same effector cells,proliferation of splenocyte was obviously higher in pGEG.gB immune mice than in pG.gB immune mice,and serum levels of IL-12 and IFN-γ increased remarkably in pG.gB and pGEG.gB immune mice.CONCLUSION Humoral and cellular immune response of mice induced by intravascular immunization of naked pDNA has an advantage in preventing and treatment the acute infection with HSV-1 in mice,and EBV plasmid vector with EBNA 1 and oriP strengthens obviously CTL activity and proliferative response.