Construction Of DNA Vaccine PRSC-gD.gC-IL-21 And Preliminary Research On Its Immunoprotection Against HSK In Mice Model

Objective:To construct DNA vaccine pRSC-gD.gC-IL-21 and to investigate their ability to induce immune response and to protect against the herpes simplex virus type1(HSV-1)infection of cornea in mice.Methods:HSV-1 genomic DNA was extracted and the gene of gC was amplified by PCR.As a template with pRSC-gD-IL-21 plasmid(in 2011,it has been building successful in our laboratory),DNA vaccine pRSC-gD.gC-IL-21 was constructed.After being identifed by molecular methods,we Package them with nanometer materials chitosan.With the method of mucosal immunization,the vaccine pRSC-gD.gC-IL-21+chitosan,the pRSC-gD-IL-21+chitosan,the blank pRSC +chitosan and the chitosan were respectively inoculated into BALB/c mice for 3 times with 2 weeks interval.A number of immunological indexes were detected 2 weeks after the last immunization.At the same time,the immunoprotective against HSV-1 challenging on cornea in mice was also evaluated.Results:By DNA sequencing,the restricted endonuclease analysis,and Western Blot expression,the construction of DNA vaccine pRSC-gD.gC-IL-21 was proved to be successful,and chitosan package rate is high.Compared with the control mice,the DNA vaccine induced the mice to generate higher levels of antibody,enhanced the splenic cell proliferation response as well as the activities of Cytotoxic T lymphocyte cell(CTL)and natural killer cell(NK),and increased the specificity slgA levels in tears.Meanwhile,the pRSC-gD.gC-IL-21+chitosan could elicit a stronger immunoprotective effect against the corneal infection of HSV-1 in mice.Conclusion:The construction of DNA vaccine pRSC-gD.gC-IL-21 was proved to be successful.Compared with the vaccine pRSC-gD-IL-21,the DNA vaccine pRSC-gD.gC-IL-21 may induce an effective immune response in immunized mice.More importantly,the efficacy against the HSV-1 challenge in mouse cornea was enhanced markedly.It can prevent the occurrence and development of the Herpes simplex keratitis(HSK).