Highly‐efficient gene transfer with retroviral vectors into human T lymphocytes on fibronectin
- 1 July 1998
- journal article
- research article
- Published by Wiley in British Journal of Haematology
- Vol. 102 (2) , 566-574
- https://doi.org/10.1046/j.1365-2141.1998.00785.x
Abstract
Genetically modified lymphocytes have been successfully used for correction of ADA deficiency in children and in controlling graft‐versus‐host disease (GvHD) after allogeneic bone marrow transplantation. Low transduction efficiencies are, however, limiting for gene therapeutic strategies based on lymphocytes. In this study we compared protocols for highly efficient gene transfer into human T cells using retroviral vector‐containing supernatant. We showed that infection of both human primary 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‐affinity nerve growth factor receptor (ΔLNGFR) as a gene transfer marker and the Herpes simplex virus thymidine kinase for negative selection. Based on ΔLNGFR expression genetically modified cells were enriched to near purity by magnetic cell sorting (MACS). Enriched cells could be shown to be highly sensitive to ganciclovir.Keywords
This publication has 22 references indexed in Scilit:
- Selective Immunoaffinity-Based Enrichment of CD34+Cells Transduced with Retroviral Vectors Containing an Intracytoplasmatically Truncated Version of the Human Low-Affinity Nerve Growth Factor Receptor (ΔLNGFR) GeneHuman Gene Therapy, 1997
- Sustained High-Level Reconstitution of the Hematopoietic System by Preselected Hematopoietic Cells Expressing a Transduced Cell-Surface AntigenHuman Gene Therapy, 1997
- HSV-TK Gene Transfer into Donor Lymphocytes for Control of Allogeneic Graft-Versus-LeukemiaScience, 1997
- Measuring gene–transfer efficiencyNature Medicine, 1996
- Cell–surface markers for assessing gene transfer into human hematopoietic cellsNature Medicine, 1996
- Colocalization of retrovirus and target cells on specific fibronectin fragments increases genetic transduction of mammalian cellsNature Medicine, 1996
- Gene Therapy for AIDS Using Retroviral Mediated Gene Transfer to Deliver HIV-1 Antisense TAR and Transdominant Rev Protein Genes to Syngeneic Lymphocytes in HIV-1 Infected Identical Twins. National Institutes of Health, Bethesda, MarylandHuman Gene Therapy, 1996
- T Lymphocyte-Directed Gene Therapy for ADA − SCID: Initial Trial Results After 4 YearsScience, 1995
- Identification and characterization of the T lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin.The Journal of cell biology, 1989
- Construction and use of a safe and efficient amphotropic packaging cell lineVirology, 1988