Efficient lentiviral transduction of Herpesvirus saimiri immortalized T cells as a model for gene therapy in primary immunodeficiencies Toscano, Miguel Frecha, Cecilia Ortega, C. Santamaría, Manuel Martín, Francisco Molina Pineda Infantas, Ignacio Jesús gene therapy vectors Lentiviral vectors T cells Heresvirus saimiri model This work was supported by V Framework European Union contract grant QLT-1999-01090 (to IJM and MS) and by Spanish Ministry of Health Grant FIS01/3143 to FM. MGT is a predoctoral fellow (FPU program) of the Spanish Ministry of Education and Culture. 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. 2025-02-01T11:56:38Z 2025-02-01T11:56:38Z 2004 journal article Toscano, M., Frecha, C., Ortega, C. et al. Efficient lentiviral transduction of Herpesvirus saimiri immortalized T cells as a model for gene therapy in primary immunodeficiencies. Gene Ther 11, 956–961 (2004). https://doi.org/10.1038/sj.gt.3302259 https://hdl.handle.net/10481/101732 10.1038/sj.gt.3302259 eng open access Springer Nature