Robust expansion of dendritic cells in vivo by hydrodynamic FLT3L-FC gene transfer

J Immunol Methods. 2014 Nov;413:69-73. doi: 10.1016/j.jim.2014.07.008. Epub 2014 Jul 24.

Abstract

Due to low numbers of endogenous dendritic cells (DCs) in vivo, exogenous DC-poietin Fms-like tyrosine kinase 3-ligand (FLT3L) is routinely used to generate DC for subsequent studies. We engineered a novel FLT3L-FC DNA construct that, when combined with hydrodynamic gene transfer (HDT), induced robust DC expansion in mice. DC generated in vivo by FLT3L-FC HDT produced cytokines in response to stimulation by an array of TLR agonists and promoted T cell proliferation. The FLT3L-FC protein produced in vivo spontaneously homodimerized to enable effective FLT signaling and the FC-domain enhanced its plasma half-life, providing an improved reagent and method to boost DC numbers.

Keywords: Cytokine; Pharmacodynamics; Pharmacokinetics; Progenitor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Count
  • Cell Proliferation
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Female
  • Gene Expression
  • Gene Transfer Techniques*
  • Half-Life
  • Hydrodynamics
  • Membrane Proteins / genetics*
  • Membrane Proteins / immunology
  • Mice
  • Mice, Inbred BALB C
  • Plasmids / metabolism*
  • Protein Multimerization
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / immunology
  • Signal Transduction

Substances

  • Membrane Proteins
  • Recombinant Fusion Proteins
  • flt3 ligand protein