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Molecular mechanism for ganciclovir resistance in human T lymphocytes transduced with retroviral vectors carrying the herpes simplex virus thymidine kinase gene.
- Source :
-
Blood [Blood] 2001 Jan 01; Vol. 97 (1), pp. 122-9. - Publication Year :
- 2001
-
Abstract
- The herpes simplex virus thymidine kinase gene type 1 (HSV-Tk) ganciclovir (GCV) system is a novel therapeutic strategy for the modulation of graft-versus-host disease (GVHD), a major complication of allogeneic stem cell transplantation (allo-SCT). Retroviral-mediated gene transfer of the HSV-Tk gene into donor T lymphocytes before allo-SCT may allow their in vivo selective depletion after treatment with GCV. The expression of the HSV-Tk gene was analyzed in vitro in CEM cells, a human lymphoblastoid cell line, transduced with 2 different vectors, each containing the HSV-Tk gene and a selectable marker gene. GCV-resistant clones were identified within the clones expressing the marker gene. Characterization of the molecular events leading to this resistance revealed a 227-bp deletion in the HSV-Tk gene due to the presence of cryptic splice donor and acceptor sites within the HSV-Tk gene sequence. Furthermore, it was confirmed that this deletion was present in human primary T cells transduced with either vector and in 12 patients who received transduced donor T cells, together with a T-cell-depleted allo-SCT. In vivo circulating transduced T cells containing the truncated HSV-Tk gene were identified in all patients immediately after infusion and up to 800 days after transplantation. In patients who received GCV as treatment for GVHD, a progressive increase in the proportion of transduced donor T cells carrying the deleted HSV-Tk gene was observed. These results suggest that the limitations within the HSV-Tk/GCV system can be improved by developing optimized retroviral vectors to ensure maximal killing of HSV-Tk-transduced cells.
- Subjects :
- Antiviral Agents administration & dosage
Antiviral Agents pharmacology
Cell Culture Techniques
Cell Line
Drug Resistance
Ganciclovir administration & dosage
Gene Expression
Genetic Therapy
Genetic Vectors pharmacology
Graft vs Host Disease therapy
Hematologic Neoplasms therapy
Humans
Lymphocyte Depletion methods
Retroviridae genetics
Sequence Analysis, DNA
T-Lymphocytes virology
Thymidine Kinase genetics
Ganciclovir pharmacology
Simplexvirus enzymology
T-Lymphocytes drug effects
T-Lymphocytes transplantation
Thymidine Kinase therapeutic use
Transduction, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 0006-4971
- Volume :
- 97
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Blood
- Publication Type :
- Academic Journal
- Accession number :
- 11133751
- Full Text :
- https://doi.org/10.1182/blood.v97.1.122