Strategies for loading dendritic cells with hepatitis C NS5a antigen and inducing protective immunity.
Dendritic cell (DC)-based vaccination strategies are promising for the treatment of cancers and infectious diseases including hepatitis C virus (HCV). As the induction of T cell-mediated immune responses by DC vaccination is highly dependent on efficient antigen loading of the DCs, the purpose of this study was to identify an optimal nonviral DC loading strategy for HCV NS5a. Furthermore, the efficacy of immunization with the NS5a-loaded DCs in comparison to plasmid encoding NS5a and NS5a protein was evaluated. Transfection of DCs with mRNA was most efficient with close to 100% of DCs expressing NS5a, whereas approximately 10% of protein-pulsed DCs and <1% of plasmid-transfected DCs expressed NS5a, suggesting remarkably different loading efficiencies. Vaccination of mice with NS5a mRNA-transfected DCs or NS5a protein-pulsed DCs resulted in significantly stronger CD4(+) and CD8(+) T-cell responses and protection from challenge with vaccinia virus expressing NS3/NS4/NS5, in comparison to vaccination with NS5a DNA-transfected DCs, plasmid encoding NS5 or rNS5a protein formulated with alum. Furthermore, vaccination with NS5a mRNA-transfected DCs was superior to vaccination with rNS5a-pulsed DCs. These data have important clinical implications, with mRNA-transfected DCs providing a safe and effective vaccination strategy against hepatitis C and possibly other pathogens.
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Vaccine and Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Babiuk
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van Drunen Littel-van den Hurk
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D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000534:Alum Compounds / Q000494:pharmacology; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects; D004279:DNA, Viral; D003713:Dendritic Cells / Q000276:immunology*; D016174:Hepacivirus / Q000276:immunology*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D012333:RNA, Messenger; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects; D014162:Transfection; D014616:Vaccinia virus / Q000276:immunology; D014761:Viral Hepatitis Vaccines / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000276:immunology* / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase
D000276:Adjuvants, Immunologic; D000534:Alum Compounds; D004279:DNA, Viral; D012333:RNA, Messenger; D014761:Viral Hepatitis Vaccines; D017361:Viral Nonstructural Proteins; C041524:aluminum sulfate; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase