HIGHLY-EFFICIENT GENE-TRANSFER WITH RETROVIRAL VECTORS INTO HUMAN T-LYMPHOCYTES ON FIBRONECTIN

Genetically modified lymphocytes have been successfully used for corre ction of ADA deficiency in children and in controlling graft-versus-ho st disease (GvHD) after allogeneic bone marrow transplantation. Low tr ansduction efficiencies are, however, limiting for gene therapeutic st rategies based on lymphocytes. In this study we compared protocols for highly efficient gene transfer into human T cells using retroviral ve ctor-containing supernatant. We showed that infection of both human pr imary T cells and CD4(+) Jurkat cells is most efficient on the matrix component fibronectin. Transduction was carried out with a retroviral vector encoding both the human intracytoplasmatically truncated low-af finity nerve growth factor receptor (Delta LNGFR) as a gene transfer m arker and the Ropes simplex virus thymidine kinase for negative select ion. Based on LNGFR expression genetically modified cells were enriche d to near purity by magnetic cell sorting (MACS). Enriched cells could be shown to be highly sensitive to ganciclovir.