High-efficiency retroviral vector mediated gene transfer into human peripheral blood CD4+ T lymphocytes

T. Rudoll, K. Phillips, S. W. Lee, S. Hull, O. Gaspar, N. Sucgang, E. Gilboa, C. Smith

Research output: Contribution to journalArticle

53 Scopus citations

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 languageEnglish (US)
Pages (from-to)695-705
Number of pages11
JournalGene Therapy
Volume3
Issue number8
StatePublished - Sep 5 1996
Externally publishedYes

Keywords

  • Human CD4 T lymphocytes
  • Retroviral vector

ASJC Scopus subject areas

  • Genetics

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    Rudoll, T., Phillips, K., Lee, S. W., Hull, S., Gaspar, O., Sucgang, N., Gilboa, E., & Smith, C. (1996). High-efficiency retroviral vector mediated gene transfer into human peripheral blood CD4+ T lymphocytes. Gene Therapy, 3(8), 695-705.