Role of TLR3 in the immunogenicity of replicon plasmid-based vaccines

Gene Ther. 2009 Mar;16(3):359-66. doi: 10.1038/gt.2008.164. Epub 2008 Dec 4.

Abstract

Replicon plasmids encoding an alphavirus RNA replicase constitute an alternative to conventional DNA plasmids with promise for DNA vaccination in humans. Replicase activity amplifies the levels of transgene mRNA through a copying process involving double-stranded (ds) RNA intermediates, which contribute to vaccine immunogenicity by activating innate antiviral responses. Toll-like receptor 3 (TLR3) is a dsRNA innate immune receptor expressed by antigen-presenting dendritic cells (DCs). Here, we test the hypothesis that TLR3 is necessary for the immunogenicity of replicon plasmid-based DNA vaccines. We show that mouse CD8 alpha(+) DC phagocytose dying replicon plasmid-transfected cells in vitro and are activated in a TLR3-dependent manner by dsRNA present within those cells. However, we find that cytotoxic T-cell responses to a replicon plasmid intramuscular vaccine are not diminished in the absence of TLR3 in vivo. Our results underscore the potential role of TLR3 in mediating immune activation by dsRNA-bearing replicon plasmid-transfected cells and indicate that other innate sensing pathways can compensate for TLR3 absence in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / immunology
  • CD8 Antigens / analysis
  • CD8-Positive T-Lymphocytes / immunology
  • Chlorocebus aethiops
  • Coculture Techniques
  • Cytotoxicity, Immunologic / immunology
  • Dendritic Cells / immunology
  • Gene Expression / immunology
  • Genes, Transgenic, Suicide
  • Genetic Vectors / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Plasmids / immunology
  • RNA, Double-Stranded / biosynthesis
  • Replicon / immunology*
  • Spleen / immunology
  • Toll-Like Receptor 3 / immunology*
  • Transfection
  • Vaccination / methods
  • Vaccines, DNA / immunology*
  • Vero Cells

Substances

  • CD8 Antigens
  • CD8 antigen, alpha chain
  • RNA, Double-Stranded
  • TLR3 protein, mouse
  • Toll-Like Receptor 3
  • Vaccines, DNA