The EGFP-tk retroviral vector, encoding enhanced green fluorescent protein (EGFP) and the herpes simplex virus thymidine kinase (HSV-tk) packaged in a Phoenix amphotropic cell line, was used to transduce healthy donor T lymphocytes. Infection yielded a mean of 41.8 +/- 9.3% SD (range 31.1-48.4%) EGFP-positive cells and a mean of 92 +/- 2% SD (range 90-94%) after cell sorting. EGFP expression remained stable for 30 d after infection. The entire gene transfer procedure had no significant effect on lymphocyte subsets and slightly reduced clonogenicity. Ganciclovir (gcv) treatment (1 microg/ml x 10 d) killed all EGFP-positive cells in the transduced and transduced/sorted populations, but had no effect on untransduced controls. Our results show that primary T lymphocytes can be transduced using an EGFP-tk vector that yields a homogeneous infected population without affecting lymphocyte subsets, function and clonogenicity.
|Titolo:||Retrovirus-mediated transfer of the herpes simplex virus thymidine kinase and enhanced green fluorescence protein genes in primary T lymphocytes|
PELICCI, PIER GIUSEPPE (Penultimo)
|Parole Chiave:||Green Fluorescent Proteins; Humans; Ganciclovir; Antiviral Agents; Simplexvirus; Luminescent Proteins; Thymidine Kinase; Transfection; Interleukin-2; Genetic Vectors; Flow Cytometry; Time Factors; Cell Division; T-Lymphocytes|
|Settore Scientifico Disciplinare:||Settore MED/04 - Patologia Generale|
|Data di pubblicazione:||set-2000|
|Digital Object Identifier (DOI):||10.1046/j.1365-2141.2000.02264.x|
|Appare nelle tipologie:||01 - Articolo su periodico|