Silencing of endogenous IL-10 in human dendritic cells leads to the generation of an improved CTL response against human melanoma associated antigenic epitope, MART-1 27-35

Clin Immunol. 2008 Mar;126(3):251-9. doi: 10.1016/j.clim.2007.11.011. Epub 2008 Jan 14.

Abstract

Dendritic cells (DC) present antigenic epitopes to and activate T cells. They also polarize the ensuing T cell response to Th1 or Th2 type response, depending on their cytokine production profile. For example, IL-12 producing DC generate Th1 type T cell response whereas IL-10 producing DC is usually tolerogenic. Different strategies--such as the use of cytokines and anti-cytokine antibodies, dominant negative forms of protein, anti-sense RNA etc.--have been employed to influence the cytokine synthetic profile of DC as well as to make DC more immunogenic. Utilizing GFP expressing recombinant adenoviruses in association with lipid-mediated transfection of siRNA, we have silenced the endogenous IL-10 gene in DC. We show that IL-10 gene silenced DC produces more IL-12 and also generates a better cytolytic T cell response against the human melanoma associated epitope, MART-1(27-35), in vitro. We also show that the GFP expressing adenoviral vector can be used to optimize the parameters for siRNA delivery in primary cells and show that RNA interference methodology can efficiently knock down virus encoded genes transcribed at very high multiplicity of infection in DC.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line, Tumor
  • Dendritic Cells / cytology
  • Dendritic Cells / metabolism*
  • Epitopes / immunology*
  • Gene Silencing*
  • Green Fluorescent Proteins
  • Humans
  • Interleukin-10 / deficiency*
  • Interleukin-10 / genetics*
  • Interleukin-10 / metabolism
  • Interleukin-12 / metabolism
  • Melanoma / immunology*
  • Melanoma / metabolism
  • Neoplasm Proteins / immunology*
  • RNA, Small Interfering
  • T-Lymphocytes, Cytotoxic / immunology*
  • Up-Regulation

Substances

  • Epitopes
  • MART-1-Melan-A(27-35) epitope
  • Neoplasm Proteins
  • RNA, Small Interfering
  • Interleukin-10
  • Green Fluorescent Proteins
  • Interleukin-12