Efficient lentiviral transduction of Herpesvirus saimiri immortalized T cells as a model for gene therapy in primary immunodeficiencies.
Infection of human T lymphocytes with the Herpesvirus saimiri (HVS) yields immortalized T-cell lines (HVS-T) which retain all the phenotypical and functional characteristics of their parental cells. This represents a new experimental model for studying genetic disorders of T lymphocytes. In spite of the efforts of many laboratories, no satisfactory way has been found so far to modify HVS-T cells genetically. We have analyzed the capacity of oncoretroviral (MLV)- and lentiviral (HIV-1)-based vectors pseudotyped with vesicular stomatitis virus glycoprotein (VSVg) to transduce HVS-T cells. HIV-1-derived vectors efficiently transduced HVS-T cell lines, reaching up to 85% of cells expressing the transgene in a single round of infection. MLV-based vectors, on the other hand, were unable to transduce more than 1% of any of the HVS-T cell lines analyzed. Lentiviral-driven gene expression was maintained constant and stable in HVS-T cells for a minimum of 48 days. We also observed that although the lentiviral transduction efficiency achieved on HVS-T cells is lower than that obtained with tumor or primary endothelial cells, it is nevertheless similar to that found with activated primary T cells.
Toscano
M G
MG
Immunology Unit, University of Granada School of Medicine, Granada, Spain.
Frecha
C
C
Ortega
C
C
Santamaría
M
M
Martín
F
F
Molina
I J
IJ
D019264:Adoptive Transfer / Q000379:methods*; D000328:Adult; D002460:Cell Line; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D015497:HIV-1 / Q000235:genetics; D006567:Herpesvirus 2, Saimiriine; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009052:Leukemia Virus, Murine / Q000235:genetics; D008297:Male; D016511:Severe Combined Immunodeficiency / Q000276:immunology / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology; D014161:Transduction, Genetic / Q000379:methods