Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

Gene Ther. 2000 Dec;7(24):2105-12. doi: 10.1038/


Dendritic cells (DC) play a key role in antigen presentation and activation of specific immunity. Much current research focuses on harnessing the potency of DC for vaccines, gene therapy, and cancer immunotherapy applications. However, DC are not readily transfected in vitro by traditional nonviral techniques. A novel DNA vaccine formulation was used to determine if DC are transfected in vitro. The formulation consists of plasmid DNA adsorbed on to cationic microparticles composed of the biodegradable polymer polylactide-co-glycolide (PLG) and the cationic surfactant, cetyltrimethylammonium bromide (CTAB). Using preparations of fluorescent-labeled plasmid DNA formulated on PLG-CTAB microparticles to study internalization by macrophages and dendritic cells in vitro and in vivo, we found that most, but not all, of the fluorescence was concentrated in endosomal compartments. Furthermore, uptake of plasmid DNA encoding HIV p55 gag adsorbed to PLG-CTAB microparticles by murine bone marrow-derived dendritic cells resulted in target gene expression, as detected by RT-PCR. The antigen was subsequently processed and presented, resulting in stimulation of an H-2kd-restricted, gag-specific T cell hybridoma. Activation of the hybridoma, detected by IL-2 production, was dose-dependent in the range of 0.1-20 microg DNA (10-2000 microg PLG) and was sustained up to 5 days after transfection. Thus, adsorption of plasmid DNA on PLG-CTAB microparticles provides a potentially useful nonviral approach for in vitro transfection of dendritic cells. Gene Therapy (2000) 7, 2105-2112.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adsorption
  • Analysis of Variance
  • Animals
  • Antigen Presentation*
  • Cetrimonium
  • Cetrimonium Compounds
  • Dendritic Cells / immunology*
  • Female
  • Gene Expression
  • Gene Products, gag*
  • Genetic Therapy / methods*
  • Hybridomas
  • Image Processing, Computer-Assisted
  • Interleukin-2 / biosynthesis
  • Lactic Acid
  • Lymphocyte Activation
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Confocal
  • Microspheres
  • Polyglycolic Acid
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polymers
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes / immunology
  • Transfection / methods*
  • Vaccines, DNA / administration & dosage*


  • Cetrimonium Compounds
  • Gene Products, gag
  • Interleukin-2
  • Polymers
  • Vaccines, DNA
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • Lactic Acid
  • Cetrimonium