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Modulating APOBEC expression enhances DNA vaccine immunogenicity.

Rafael Ribeiro Almeida1,2, Rui André Saraiva Raposo3, Fernanda Caroline Coirada1

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DNA vaccine immunogenicity in humans is limited by APOBEC-mediated plasmid degradation. Silencing APOBEC2 with a DNA vaccine enhanced immune responses, offering a strategy to improve vaccine effectiveness.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Vaccinology

Background:

  • DNA vaccines face challenges in eliciting robust immune responses in humans.
  • Mechanisms of double-stranded DNA (dsDNA) sensing significantly influence DNA vaccine immunogenicity.
  • APOBEC (apolipoprotein B mRNA-editing, catalytic polypeptide) enzymes are implicated in nucleic acid metabolism and immune regulation.

Purpose of the Study:

  • To investigate the hypothesis that APOBEC-mediated plasmid degradation suppresses DNA vaccine immunogenicity in humans.
  • To elucidate the role of STING (stimulator of interferon genes) and TBK-1 (TANK-binding kinase 1) signaling in APOBEC upregulation.
  • To develop a strategy to enhance DNA vaccine efficacy by targeting APOBEC activity.

Main Methods:

  • Investigated dsDNA sensing pathways involving STING and TBK-1, leading to IFN-β induction.
  • Examined the effect of APOBEC3A mRNA upregulation on DNA vaccine components.
  • Assessed the impact of murine APOBEC2 expression on plasmid levels and expression in HEK293T cells.
  • Developed and tested a bicistronic DNA vaccine encoding an immunogen and an APOBEC2-specific shRNA (short hairpin RNA) in vitro and in vivo.

Main Results:

  • STING and TBK-1 sensing induced IFN-β, upregulating APOBEC3A via protein kinase C signaling.
  • APOBEC2 expression significantly reduced intracellular plasmid levels, plasmid-encoded mRNA, and reporter gene expression.
  • The bicistronic DNA vaccine effectively silenced APOBEC2, leading to increased frequency of IFN-γ-secreting T cells.

Conclusions:

  • APOBEC-mediated plasmid degradation is a key mechanism suppressing DNA vaccine immunogenicity in humans.
  • Targeting APOBEC activity, specifically APOBEC2, through RNA interference can enhance DNA vaccine-induced cellular immune responses.
  • These findings provide novel insights into dsDNA sensing pathways and offer a promising strategy for improving human DNA vaccine efficacy.