Immunity and protection by adoptive transfer of dendritic cells transfected with hepatitis C NS3/4A mRNA.
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.
Yu
Hong
H
Vaccine and Infectious Disease Organization, University of Saskatchewan, 120 Veterinary Road, Saskatoon, Sask. S7N 5E3, Canada.
Babiuk
Lorne A
LA
van Drunen Littel-van den Hurk
Sylvia
S
D019264:Adoptive Transfer / Q000379:methods*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology* / Q000821:virology; D004195:Disease Models, Animal; D018274:Electroporation; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology; D006526:Hepatitis C / Q000276:immunology* / Q000517:prevention & control*; D047908:Intracellular Signaling Peptides and Proteins; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection; D014611:Vaccination / Q000379:methods*; D014615:Vaccinia / Q000517:prevention & control; D014616:Vaccinia virus / Q000235:genetics / Q000254:growth & development; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology; D014766:Viremia / Q000517:prevention & control; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D002352:Carrier Proteins; D049452:Green Fluorescent Proteins; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C455041:NS4A cofactor peptide, Hepatitis C virus; C485184:enhanced green fluorescent protein