T cell transduction and suicide with an enhanced mutant thymidine kinase.
Retroviral transfer of Herpes simplex virus thymidine kinase to T cells has been used to confer sensitivity to the antiviral agent ganciclovir. This has allowed therapeutic approaches to be developed in which T cells mediating graft-versus-host disease after bone marrow transplantation can be selectively eliminated by the administration of ganciclovir. Although the strategy has been shown to be generally successful in early clinical trials, there are concerns about possible resistance to ganciclovir and the risk of myelosuppressive side-effects at the doses required to induce T cell suicide. We have incorporated the enhanced mutant HSV-TKSR39 into retroviral vectors tailored to exhibit high levels of expression in T cells and have used protocols optimized for the transduction and selection of primary lymphocytes. We demonstrate that leukemic and primary T cells can be efficiently transduced and highly enriched under conditions that should be readily adaptable for clinical use. T cells carrying HSV-TKSR39 were inhibited by exposure to ganciclovir at concentrations an order of magnitude below those required for wild-type HSV-TK. The less toxic agent aciclovir also eliminated T cells transduced with HSV-TKSR39 (but not HSV-TK), underlining the increased therapeutic potential of the mutant suicide gene system in the bone marrow transplantation setting.
Qasim
W
W
Molecular Immunology Unit, Institute of Child Health, University College London, UK.
Thrasher
A J
AJ
Buddle
J
J
Kinnon
C
C
Black
M E
ME
Gaspar
H B
HB
D000212:Acyclovir / Q000627:therapeutic use; D000998:Antiviral Agents / Q000627:therapeutic use; D016026:Bone Marrow Transplantation / Q000276:immunology*; D002460:Cell Line; D002478:Cells, Cultured; D015774:Ganciclovir / Q000627:therapeutic use; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D006086:Graft vs Host Disease / Q000628:therapy*; D006801:Humans; D015458:Leukemia, T-Cell; D009154:Mutation; D012190:Retroviridae / Q000235:genetics; D018139:Simplexvirus / Q000201:enzymology*; D013601:T-Lymphocytes / Q000821:virology*; D013937:Thymidine Kinase / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods
D000998:Antiviral Agents; D013937:Thymidine Kinase; D015774:Ganciclovir; D000212:Acyclovir