Hematological reconstitution and gene therapy: retroviral transfer of the bacterial beta-galactosidase activity into human hematopoietic CD34+ cell populations and into T lymphocytes derived from the peripheral blood.
We report the possibility to transfer marker genes coding for beta-galactosidase activity using retroviral vectors into human peripheral blood CD34+ cells, peripheral blood T-lymphocytes and into the growth factor-dependent human hematopoietic cell line TF-1. Using the MFG-nisLacZ and the FLac vector and various packaging cell lines, we demonstrated retroviral transfer and high expression of a bacterial beta-galactosidase activity induced by the nisLacZ gene or the Sh-ble/LacZ gene. Kinetics of expression of the transgenes were analyzed both in primary cells and cell lines. Absence of cytotoxicity related to the expression of the bacterial beta-galactosidase was assessed in both cell types. These results open interesting prospectives for the use of the beta-galactosidase activity to mark and follow the fate of genetically modified cells isolated from patients prior to reimplantation.
D015703:Antigens, CD; D018952:Antigens, CD34; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D015316:Genetic Therapy / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology / Q000473:pathology; D006801:Humans; D009369:Neoplasms / Q000097:blood / Q000276:immunology; D012190:Retroviridae; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D015703:Antigens, CD; D018952:Antigens, CD34; D001616:beta-Galactosidase