Immunity and protection by adoptive transfer of dendritic cells transfected with hepatitis C NS3/4A mRNA

Abstract

In this study, we tested the hypothesis that adoptive transfer of dendritic cells (DCs) transfected ex vivo with mRNA encoding hepatitis C virus (HCV) NS3/4A would initiate potent HCV-specific protective immune responses in vivo. Murine DCs were transfected with NS3/4A mRNA or eGFP mRNA using either electroporation or Transmessenger Transfection Reagent and then used for adoptive transfer. Electroporation resulted in higher transfection efficiency but lower levels of eGFP and NS3/4A expression when compared to transfection with Transmessenger. The murine NS3/4A mRNA-transfected DCs were functional in T cell activation in vitro. Adoptive transfer of NS3/4A mRNA-transfected DCs resulted in migration to regional lymph nodes, strong cellular immune responses and protection from challenge with vaccinia virus expressing NS3/NS4/NS5 in mice. Furthermore, although Transmessenger mediated transfection was less efficient than electroporation in terms of number of transfected cells, the DCs transfected with NS3/4A mRNA and Transmessenger expressed higher levels of protein and induced stronger immune responses and protection than DCs transfected with NS3/4A mRNA by electroporation. Since no study has explored the in vivo efficacy of mRNA-transfected DC-mediated vaccination against viral diseases, including hepatitis C, our study provided a novel vaccination strategy against hepatitis C as well as other pathogens.