Preparation Of HSV-2 GD Antigen With GB CTL Epitoeps And Its Immune Effect Evaluation

HSV-2 ( Herpes simplex virus 2 ) is a double-stranded DNA virus, it can cause genital herpes. HSV-2 infection is prevalent in the global, and is severer in developing countries. HSV-2 can also increase the probability of AIDS infection and cervical cancer. The vaccine plays an important role in the prevention and therapy of HSV-2 infection. The current vaccine development works focus on HSV-2 glycoproteins.Eleven kinds of glycoproteins on HSV-2 surface are the main immunogens, glycoprotein D ( gD ) and glycoprotein B ( gB ) are the most important immunogens. gD is of high homology and conservation, gB could induce neutralizing antibody and cytotoxic T cell responses. CTL can bind with MHCâ… molecule to stimulate cellular immunity, which is an important component of the immune responses against HSV-2. In this study, the recombinant gD-gBCTL gene was constructed, which contains HSV-2 gD and two gB CTL epitopes. gD-gBCTL protein was expressed in both baculovirus expression system and the prokaryotic expression system in order to evaluate its immune effects.In the baculovirus expression system, gD gene and gD-gBCTL gene were both cloned into pFastBac1 vector to construct recombinant plasmids, then were transformed into DH10BacTM to generate the recombinant bacmids. The recombinant bacmids transfected Sf9 cells, the recombinant proteins were expressed. After optimizing expression conditions, gD and gD-gBCTL proteins were both expressed in high levels, which laid the foundation for further purification and the immune effect evaluation.In the prokaryotic expression system, pET-22b(+)-gD-gBCTL was constructed to transform E.coli BL21 (DE3). The recombinant protein was highly expressed after optimizing the expressing conditions. Then the purifed protein was used to immunize Balb/c mice with different adjuvants. Immune effects were preliminary evaluated, then mice were challenged with HSV-2 strain 333. The results indicated this recombinant protein could induce protective immunity against genital herpes in mice.