Construction of a recombinant eukaryotic expression vector containing HSV 2gD and its immune responses

AIM: To construct and express pcDNA3.1-IL-18-HSV P6 recombinant DNA vaccine,and to observe the immune responses of pcDNA3.1-IL-18-HSV P6 genetic vaccine.METHODS: Genes of P6 and IL-18 were cloned into the eukaryotic expression vector pcDNA3.1(-) respectively,and confirmed by enzyme digestion and sequence analysis,the recombinant plasmid pcDNA3.1-IL-18-HSVP6 was transformed into CHO cells.The expressed protein was characterized by indirect immunofluorescence and Western blotting.The recombinant plasmid pcDNA3.1-IL-18-HSVP6 was used to inoculate BALB/c mice by intramuscular injection for three times,once a week.One week after the last vaccination,the levels of specific IgG antibody,IFN-γ and IL-18 were detected by ELISA;One month after the last vaccination,spleen cells of vaccinated mice were separated and proliferation of spleen lymphocytes were detected by MTT assay.RESULTS: Recombinant pcDNA3.1-IL-18-HSV P6 plasmid was confirmed by enzyme digestion and DNA sequencing.After inoculated by pcDNA3.1-IL-18-HSV P6 vaccine,the mice could produce higher level of splenocytes proliferation and secrected higher level of IFN-γ,IL-18 and specific antibody.CONCLUSION: DNA vaccine of pcDNA3.1-IL-18-HSV P6 has been successfully constructed,and it can effectively induce humoral and cellular immune responses,which prorided a basis for constructing new type of DNA vaccine.