Abstract
Genetic modification of peripheral blood T lymphocytes been proposed as a therapeutic strategy for treating congenital disorders, cancer and viral diseases. Central to all T lymphocyte-based gene therapy strategies is the ability to efficiently and stably deliver genes into primary T lymphocytes. In this study, we sought to increase the gene transfer efficiency in CD4+ peripheral blood T lymphocytes using procedures which could be utilized in clinical applications. In order to quantify the gene transfer efficiency in primary CD4+ T cells, a high-titer retroviral vector which efficiently expresses a truncated version of the human low-affinity nerve growth factor receptor (ΔLNGFR) was constructed. Transduced cells were then accurately enumerated with immunofluorescence staining and fluorescence activated cell sorting (FACS) analysis. Using this system, a supernatant-based gene transfer procedure was developed which routinely yields gene transfer efficiencies of 25-40% into a wide repertoire of both freshly obtained and cryopreserved peripheral blood CD4+ T lymphocytes.
Original language | English (US) |
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Pages (from-to) | 695-705 |
Number of pages | 11 |
Journal | Gene Therapy |
Volume | 3 |
Issue number | 8 |
State | Published - 1996 |
Externally published | Yes |
Keywords
- Human CD4 T lymphocytes
- Retroviral vector
ASJC Scopus subject areas
- Genetics