Potency of mature CD40L RNA electroporated dendritic cells correlates with IL-12 secretion by tracking multifunctional CD8(+)/CD28(+) cytotoxic T-cell responses in vitro

J Immunother. 2011 Jan;34(1):45-57. doi: 10.1097/CJI.0b013e3181fb651a.


Electroporation of mature dendritic cells (DC) with RNA-encoding CD40L greatly enhances the production of interleukin (IL)-12, a proinflammatory cytokine necessary for the induction of T-cell immunity. Results presented herein reveal a correlation between the priming of CD28(+) antigen-reactive effector memory cytotoxic T lymphocytes (CTL) displaying 3 or 4 simultaneous effector functions and the quantity of IL-12 produced by postmaturation electroporation-CD40L DC. By using multiparameter flow cytometry, the quantities of IL-12 needed to prime naive antigen-reactive T cells to simultaneously produce interferon-γ and tumor necrosis factor-α in the presence or absence of IL-2 secretion in conjunction with lytic activity defined by CD107a expression can be used to determine the overall potency of a DC product. In the presence of IL-12, CTL differentiation toward lytic function is not accompanied by a reduction in the secretion of interferon-γ and tumor necrosis factor-α. Therefore, by measuring the availability of IL-12 one can predict the potency of a DC immunotherapeutics in relation to its ability to drive distinct effector memory CTL subsets with multifunctional activities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD28 Antigens / biosynthesis
  • CD40 Ligand / genetics*
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism*
  • Electroporation*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Humans
  • Interferon-gamma / biosynthesis
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / metabolism*
  • Interleukin-2 / immunology
  • Interleukin-2 / metabolism
  • Lymphocyte Activation
  • Lysosomal-Associated Membrane Protein 1 / genetics
  • RNA / genetics*
  • Signal Transduction
  • T-Lymphocytes, Cytotoxic / immunology*
  • Tumor Necrosis Factor-alpha / biosynthesis


  • CD28 Antigens
  • Interleukin-2
  • Lysosomal-Associated Membrane Protein 1
  • Tumor Necrosis Factor-alpha
  • CD40 Ligand
  • Interleukin-12
  • RNA
  • Interferon-gamma