Highly efficient retroviral gene transfer into human primary T lymphocytes derived from peripheral blood.
T lymphocytes are a promising cell vehicle for gene therapy purposes. By cocultivating retroviral vector producing cells and target cells, highly efficient gene transfer was achieved with activated human T lymphocytes derived from peripheral blood with vectors carrying different forms of the bacterial beta-galactosidase gene including the regular LacZ gene, the Sh-ble::LacZ gene and the nlsLacZ gene. Infection kinetics of T cells activated by a combination of monoclonal antibodies directed against CD2 and CD28 indicated that the highest efficiencies of transduction were obtained when the cocultivation began 4 days after stimulation. In fact, with the FLac vector, a new retroviral vector which expresses the Sh-ble::LacZ gene, we observed up to 78% transduction efficiency assessed by X-gal staining performed 2 days after the end of the cocultivation. Expression of the transduced genes was observed throughout the period of culture. Neither the cocultivation step nor the expression of the transduced Sh-ble::LacZ gene altered cell culture proliferation or the expression of selected cell surface antigens. In addition, we showed that CD4+ and CD8+ T cells were equally transduced.
Imbert
A M
AM
Institut Paoli-Calmettes, Marseille, France.
Costello
R
R
Imbert
J
J
Mannoni
P
P
Bagnis
C
C
D000911:Antibodies, Monoclonal / Q000276:immunology; D018801:CD2 Antigens / Q000276:immunology; D018106:CD28 Antigens / Q000276:immunology; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016130:Immunophenotyping; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D014162:Transfection
D000911:Antibodies, Monoclonal; D018801:CD2 Antigens; D018106:CD28 Antigens; D004274:DNA, Recombinant