Gene transfer into stimulated and unstimulated T lymphocytes by HIV-1-derived lentiviral vectors

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Serval ID
serval:BIB_7BF1A29E909C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Gene transfer into stimulated and unstimulated T lymphocytes by HIV-1-derived lentiviral vectors
Journal
Gene Therapy
Author(s)
Costello E., Munoz M., Buetti E., Meylan P. R., Diggelmann H., Thali M.
ISSN
0969-7128 (Print)
Publication state
Published
Issued date
04/2000
Volume
7
Number
7
Pages
596-604
Notes
Journal Article Research Support, Non-U.S. Gov't --- Old month value: Apr
Abstract
Genetic modification of T lymphocytes holds great potential for treatments of cancer, T cell disorders and AIDS. While in the past recombinant murine retroviruses were the vectors of choice for gene delivery to T cells, vectors based on lentiviruses can provide additional benefits. Here, we show that VSV-G pseudotyped HIV 1 vector particles delivering the enhanced green fluorescent protein (EGFP) efficiently transduce human T lymphocytes. Transduction efficiency was optimal when infection included centrifugation of cells with concentrated vector supernatant in the presence of Polybrene. In contrast to previous reports describing murine retrovirus-mediated gene transfer to T lymphocytes, fibronectin did not improve the transduction efficiency of the VSVG-pseudotyped HIV-1 particles. Similar gene transfer efficiencies were observed following stimulation of cells with PHA/IL-2 or anti-CD3i/CD28i antibodies, although greater transgene expression was observed in the latter case. Interestingly, production of vectors in the absence of the accessory proteins Vif, Vpr, Vpu and Nef was accompanied by a 50% decrease in transduction efficiency in activated T cells. Transduction of T cells that were not stimulated before infection was achieved. No transduction of non-prestimulated cells was observed with a GAL V-pseudotyped murine retroviral vector. The requirement for accessory proteins in non-prestimulated cells was more pronounced. Our results have implications for lentiviral vector targeting of other cells of the hematopoietic system including stem cells.
Keywords
Cell Line Gene Expression Gene Therapy/*methods Genetic Engineering/methods Genetic Vectors/*therapeutic use Green Fluorescent Proteins HIV-1/*genetics Humans Indicators and Reagents Luminescent Proteins/genetics Lymphocyte Activation/*immunology T-Lymphocytes/*immunology Transfection/*methods
Pubmed
Web of science
Open Access
Yes
Create date
25/01/2008 14:25
Last modification date
20/08/2019 14:37
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