Modification of tumor cells with Fas (CD95) antigen gene and Fas ligand (CD95L) gene transfection by electroporation for immunotherapy of cancer

Mol Biotechnol. 2003 Sep;25(1):79-87. doi: 10.1385/MB:25:1:79.

Abstract

Electroporation is a method for introducing DNA into cells by using a high-voltage electric field. This method is very simple and easily manipulated. We describe here a method for the modification of tumor cells with the Fas/Apo-1 (CD95) antigen-gene and Fas ligand (FasL)-gene transfection through the use of electroporation, and suggest that the Fas-FasL system is a good target for the induction of apoptosis-mediated antitumor activity. The Fas receptor/ligand system induces apoptosis and plays an important role in regulation of the immune system. In the method described, hepatoma MH134 (Fas- and FasL-) is transfected with murine Fas and FasL cDNA. A single administration of monoclonal anti-Fas antibody efficiently suppresses the growth of F6b (MH134+Neo+Fas) tumors but not that of N1d (MH134+Neo) tumors in gld/gld lpr/lpr mice. MH134+Neo+FasL tumor cells were rejected after the induction of inflammation with infiltration of neutrophils in mice. These results suggest that electroporation and Fas-mediated apoptosis are a good method for inducing of antitumor activity.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacology
  • Cell Line, Tumor / immunology
  • Cell Line, Tumor / physiology
  • Cell Survival / drug effects
  • DNA, Complementary / genetics
  • Dose-Response Relationship, Drug
  • Drug Screening Assays, Antitumor / methods
  • Electroporation
  • Fas Ligand Protein
  • Female
  • Flow Cytometry / methods
  • Gene Expression / genetics
  • Gentamicins / pharmacology
  • Immunotherapy / methods*
  • Inbreeding
  • Injections, Intradermal
  • Male
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / immunology
  • Mice
  • Mice, Inbred C3H
  • Neoplasms / genetics
  • Neoplasms / immunology
  • Neoplasms / therapy*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection / methods
  • fas Receptor / genetics*
  • fas Receptor / immunology

Substances

  • Antibodies, Monoclonal
  • DNA, Complementary
  • Fas Ligand Protein
  • Fasl protein, mouse
  • Gentamicins
  • Membrane Glycoproteins
  • fas Receptor
  • antibiotic G 418