Expression of HSV-TK suicide gene in primary T lymphocytes: The dog as a preclinical model

E. M. Weissinger, M. Franz, C. Voss, C. Bonini, E. Kremmer, H. J. Kolb

Research output: Contribution to journalArticlepeer-review

Abstract

Expression of suicide genes (e.g. herpes simplex virus thymidine kinase, HSV-TK) in T cells is an appealing approach to regulate graft-versus-host disease in adoptive immunotherapy. Here we report the optimization of retroviral infection of canine T cells. Canine T cells were stimulated either with phytohemagglutinin (PHA, 2 μg/ml) for 24-72 hours or with 100 U/ml interleukin-2 for seven days. Stimulated cells were co-cultivated with irradiated virus-producing cells. Transduction efficiencies ranged from 4% to 45% using PG13, a gibbon ape leukemia virus envelope (env) pseudotyped packaging cell line. Infection of cells with GPenvAM12, expressing the amphotropic Moloney routine leukemia virus env, did not yield a satisfactory percentage of transduced cells. Enrichment of transduced cells was performed using immunoselection, and gave a purity of up to 98%. Transfusion of 1 10 6 transduced cells per kilogram body weight showed that transduced cells could convert mixed chimerism to 100% and transfer immunity to a specific antigen. Transduced cells were repeatedly detected in peripheral blood and bone marrow by polymerase chain reaction with primers specific for the HSV-TK gene. We have demonstrated the feasibility of using the canine model to study gene therapy as a preclinical model.

Original languageEnglish
Pages (from-to)25-33
Number of pages9
JournalCytokines, Cellular and Molecular Therapy
Volume6
Issue number1
Publication statusPublished - 2000

Keywords

  • Canine model
  • Donor leukocyte transfusion
  • Gene therapy
  • Hematopoietic stem cell transplantation
  • Suicide gene (e.g. HSV-TK)

ASJC Scopus subject areas

  • Pharmacology
  • Immunology and Allergy
  • Immunology

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