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  4. Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors
 
research article

Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors

Kvell, Krisztian
•
Nguyen, Tuan H
•
Salmon, Patrick
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2005
Molecular Therapy

Recently, using HIV-1-derived lentivectors, we obtained efficient transduction of primary human B lymphocytes cocultured with murine EL-4 B5 thymoma cells, but not of isolated B cells activated by CD40 ligation. Coculture with a cell line is problematic for gene therapy applications or study of gene functions. We have now found that transduction of B cells in a system using CpG DNA was comparable to that in the EL-4 B5 system. A monocistronic vector with a CMV promoter gave 32 +/- 4.7% green fluorescent protein (GFP)(+) cells. A bicistronic vector, encoding IL-4 and GFP in the first and second cistrons, respectively, gave 14.2 +/- 2.1% GFP(+) cells and IL-4 secretion of 1.3 +/- 0.2 ng/10(5) B cells/24 h. This was similar to results obtained in CD34(+) cells using the elongation factor-1alpha promoter. Activated memory and naive B cells were transducible. After transduction with a bicistronic vector encoding a viral FLIP molecule, vFLIP was detectable by FACS or Western blot in GFP(+), but not in GFP(-), B cells, and 57% of sorted GFP(+) B cells were protected against Fas ligand-induced cell death. This system should be useful for gene function research in primary B cells and development of gene therapies.

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Type
research article
DOI
10.1016/j.ymthe.2005.05.010
Author(s)
Kvell, Krisztian
Nguyen, Tuan H
Salmon, Patrick
Glauser, Frédéric
Werner-Favre, Christiane
Barnet, Marc
Schneider, Pascal
Trono, Didier  
Zubler, Rudolf H
Date Issued

2005

Published in
Molecular Therapy
Volume

12

Start page

892

End page

899

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LVG  
Available on Infoscience
September 5, 2005
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/215905
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