Kinetics of cell death in T lymphocytes genetically modified with two novel suicide fusion genes

Gene Ther. 2003 Jul;10(14):1189-97. doi: 10.1038/sj.gt.3301977.

Abstract

Donor lymphocyte infusions (DLI) following allogeneic stem cell transplantation are known to mediate graft-versus-leukemia effect (GVL). A major side effect of these immunotherapies is the development of graft-versus-host diseases (GVHD). One promising approach to prevent GVHD is to genetically modify donor T cells with a suicide mechanism that can be induced in the case of GVHD. Here we report on a retroviral vector containing the death effector domain (DED) of the human Fas-associated protein with death domain (FADD). The DED was fused to two copies of an FKBP506-binding protein and a truncated version of the human low-affinity receptor for nerve growth factor (LNGFR). Activation of the death signal pathway can be triggered upon the addition of chemical inducers of dimerization. This construct was functionally compared to an optimized HSV-TK vector in which a hypersensitive mutant of the herpes simplex virus thymidine kinase gene (TK39) was fused to a cytoplasmic truncated version of the cell surface antigen CD34. A direct comparison between both vectors in primary T lymphocytes showed that the number of T cells transduced with vectors containing the DED was significantly reduced within 24 h of drug administration whereas ganciclovir treatment of TK39-transduced T cells showed a delay in cell death of approximately 3-4 days. Our results indicate that constructs containing the DED may prove to be the most efficient mechanism to quickly eliminate alloreactive T cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / therapeutic use
  • Cell Death / genetics
  • Coenzyme A Ligases / genetics*
  • Escherichia coli Proteins / genetics*
  • Ganciclovir / therapeutic use
  • Genetic Engineering
  • Genetic Therapy / methods*
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Graft vs Host Disease / etiology
  • Graft vs Host Disease / immunology
  • Graft vs Host Disease / therapy*
  • Humans
  • Lymphocyte Transfusion / adverse effects*
  • Receptors, Nerve Growth Factor / genetics
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / genetics
  • Retroviridae / genetics
  • Simplexvirus / enzymology
  • T-Lymphocytes / metabolism*
  • T-Lymphocytes / pathology
  • Tacrolimus Binding Proteins / genetics
  • Thymidine Kinase / genetics

Substances

  • Antiviral Agents
  • Escherichia coli Proteins
  • Receptors, Nerve Growth Factor
  • Recombinant Fusion Proteins
  • Thymidine Kinase
  • Tacrolimus Binding Proteins
  • Coenzyme A Ligases
  • acyl-coenzyme A synthetase, E coli
  • Ganciclovir