Naked Gene Vaccine Of Encoding Glycoprotein B Induces Humoral And Cellular Immunity Of HSV-Ⅰ Infection In Mice

Objectives: Transfecting the mice by injecting pDNA which encoded as HSV-1 gB through tail vein to testify efficiency of high-effect transmitting system on gene vaccine treating infectious diseases and analyze antiviral immune response induced by gene vaccine.Methodology:Injecting normal saline with different dosages and different pDNAs to Balb/c mice and immune again a week later. Observe different survival rates of experimental and matched group animals being implemented immunization by different combination of pDNAs on infecting fatal dosage of HSV-I. This paper evaluates its immunological effect through detecting latent infection, determining cytokine concentrations of serum, testing cytotoxic reaction and neutralization test.Results:1. Survival rate of 5μg pGgB immune group improves remarkably (P<0.01) comparing to the matched group among mice inoculated by HSV-1 gB expression plasmid (pG.gB); and there's great difference (P<0.05) among survival rates of 5μg immune group and 3μg and 1μg pGgB immune group.2. Survival rate of 5μg pGgB immune group improves remarkably (P<0.01) comparing to the matched group among mice inoculated by HSV-1 gB expression plasmid (pGEGgB) with EBNA1 gene and orip sequence; and there's great difference (P<0.05) among survival rates of 5μg immune group and 3μg and 1μg pGgB immune group but no significant difference between survival rates of 5μg pG.gB immune group and pGgB immune group.3. Potency of neutralizing antibody generated by induction of 5μg pGgB or pGEG.gB owns significant difference (P<0.05) from the matched group. Groups received 1μg pGgB or pGEG.gB also show inhibiting effect toward formation of HSV-1 plaque, but the inhibitory rate is much lower than that of the 5μg pDNAs group.4.5μg pGEGgB immune group owns great cytotoxicity (P<0.05) on P815.gB cells comparing to the same effector cells, and we have not seen stronger cytotoxic effect of 1μg pGEGgB immune mice on P815.gB. Proliferation of pGEG.gB-immune mice splenocyte is obviously higher than pG.gB immune mice according to prolifereation experiment of splenocyte (P<0.05).5. Analyzing by ELISA, IL-12 and IFN-y concentrations of serum of pGgB and pGEG.gB immune mice increase remarkably (P<0.05) comparing to the matched groups. But there is no remarkable difference among IL-12 and IFN-y concentrations of experimental groups with different dosages.Conclusion:The hydrodynamics-based procedure is a convenient and efficient gene transfer method for gB gene in vivo; Intravascular immunization of Naked pDNA is able to induce humoral and cellular immune response which indicates that it has an advantage in preventing ande treatment mice from HSV-1 acute infection; EBV plasmid vector with EBNA1 and oriP strengthens CTL activity and proliferation obviously.