Lipoplexes carrying mRNA encoding Gag protein modulate dendritic cells to stimulate HIV-specific immune responses

Nanomedicine (Lond). 2013 Jan;8(1):77-87. doi: 10.2217/nnm.12.97. Epub 2012 Aug 14.

Abstract

Aim: Cationic lipids (Lipofectamine™ [Invitrogen, Merelbeke, Belgium] and 1,2-dioleoyl-3-trimethylammonium-propane/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and polymers (jetPEI™ and in vivo-jetPEI™ [Polyplus-transfection, Illkirch, France]) were evaluated for their potential to deliver mRNA to monocyte-derived dendritic cells.

Materials & methods: Lipoplexes and polyplexes, containing mRNA encoding GFP or Gag protein, were incubated with human monocyte-derived dendritic cells and transfection efficiencies were assessed by flow cytometry.

Results: Lipofectamine was by far the most efficient in mRNA delivery, therefore it was used in further experiments. Incubation of monocyte-derived dendritic cells isolated from HIV-1-positive donors with mRNA encoding Gag protein complexed to Lipofectamine resulted in 50% transfection. Importantly, coculture of these Gag-transfected dendritic cells with autologous T cells induced an over tenfold expansion of IFN-γ- and IL-2-secreting CD4(+) and CD8(+) T cells.

Conclusion: Cationic lipid-mediated mRNA delivery may be a useful tool for therapeutic vaccination against HIV-1. This approach can be applied to develop vaccination strategies for other infectious diseases and cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dendritic Cells / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Products, gag / genetics*
  • HIV / immunology*
  • Humans
  • Immunophenotyping
  • Microscopy, Fluorescence
  • RNA, Messenger / genetics*

Substances

  • Gene Products, gag
  • RNA, Messenger