Lentiviral-mediated gene transfer into human lymphocytes: role of HIV-1 accessory proteins

Details

Serval ID
serval:BIB_1E8925911DE1
Type
Article: article from journal or magazin.
Collection
Publications
Title
Lentiviral-mediated gene transfer into human lymphocytes: role of HIV-1 accessory proteins
Journal
Blood
Author(s)
Chinnasamy D., Chinnasamy N., Enriquez M. J., Otsu M., Morgan R. A., Candotti F.
ISSN
0006-4971 (Print)
ISSN-L
0006-4971
Publication state
Published
Issued date
2000
Volume
96
Number
4
Pages
1309-16
Language
english
Notes
Chinnasamy, D
Chinnasamy, N
Enriquez, M J
Otsu, M
Morgan, R A
Candotti, F
eng
Blood. 2000 Aug 15;96(4):1309-16.
Abstract
Resting lymphocytes are refractory to gene transfer using Moloney murine leukemia virus (MMLV)-based retroviral vectors because of their quiescent status. Recently, it has been shown that lentiviral vectors are capable of transferring genes into nondividing and terminally differentiated cells. We used human immunodeficiency virus type-1 (HIV-1)-based vectors expressing enhanced green fluorescent protein (EGFP) driven by different promoters (CMV, MPSV, or PGK) and investigated their ability to transduce human T- and B-cell lines, as well as resting or activated primary peripheral and umbilical cord blood lymphocytes. The effects of the presence or the absence of HIV-1 accessory proteins (Vif, Vpr, Vpu, and Nef) in the vector system were also assessed. Flow cytometry analysis showed no differences in the ability of these vectors of transferring the reporter gene into lymphocytic lines and mitogen-stimulated primary lymphocytes in the presence or the absence of HIV-1 accessory proteins (APs). Similarly, viral supernatants generated in the presence of accessory genes could efficiently transduce various subsets of resting lymphocytes and provide long-term expression of the transgene. No significant transduction-induced changes in cell activation or cycling status were observed and Alu-HIV-1 long terminal repeat polymerase chain reaction (LTR PCR) analysis demonstrated integration of the vector sequences at the molecular level. In contrast, in the absence of HIV-1 APs, lentiviral vectors failed to integrate and express the transgene in resting lymphocytes. These results show that transduction of primary resting lymphocytes with HIV-1-based vectors requires the presence of viral accessory proteins. (Blood. 2000;96:1309-1316)
Keywords
*B-Lymphocytes, Cell Line, Gene Expression, *Gene Transfer Techniques, *Genetic Therapy, *Genetic Vectors, Green Fluorescent Proteins, HIV-1/*genetics, Humans, *Lentivirus, Luminescent Proteins/genetics, Lymphocyte Activation, Promoter Regions, Genetic, *T-Lymphocytes, Viral Proteins/genetics
Pubmed
Create date
01/11/2017 10:29
Last modification date
20/08/2019 12:54
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