|
|
Prepublished online as a Blood First Edition Paper on October 24, 2002; DOI 10.1182/blood-2001-12-0249.
Submitted December 14, 2001
Accepted October 7, 2002
Efficient transduction of normal human B lymphocytes and non-dividing myeloma B cells with HIV-1-derived lentiviral vectors
Fabrice Bovia, Patrick Salmon, Thomas Matthes, Krisztian Kvell, Tuan H Nguyen, Christiane Werner-Favre, Marc Barnet, Monika Nagy, Florence Leuba, Jean-Francois Arrighi, Vincent Piguet, Didier Trono, and Rudolf H Zubler*
Division of Hematology, Department of Medicine, University Hospital, Geneva, Switzerland
Department of Genetics and Microbiology, University Hospital, Geneva, Switzerland
Department of Dermatology, University Hospital, Geneva, Switzerland
* Corresponding author; email: rudolf.zubler{at}hcuge.ch.
We studied the transduction of normal human B lymphocytes and myeloma cells with lentiviral vectors. In peripheral blood B cells which had been activated with helper T cells (murine thymoma EL-4 B5) and cytokines, multiply attenuated HIV-1-derived vectors pseudotyped with VSV G envelope protein achieved expression of green fluorescent protein (GFP) in 27 ± 12% (mean ± 1 SD, median = 27%) of B cells in different experiments. When compared in parallel cultures, the transducibility of B cells from different donors exhibited very little variation. The human CMV promoter gave 4- to 6-fold higher GFP expression than did the human elongation factor-1 promoter. A murine retroviral vector pseudotyped with VSV-G protein proved inefficient even in mitotically active normal B cells. B cells freshly stimulated with Epstein-Barr virus were also transducible by HIV vectors (24 ± 9%), but B cells activated with CD40 ligand and cytokines resisted transduction. Thus, different culture systems gave very different results. Freshly isolated, non-dividing myeloma cells were efficiently transduced by HIV vectors; for six myelomas the range was 14 to 77% (median of 28%) GFP+ cells. HIV vectors with a mutant integrase led to no significant GFP signal in normal B or myeloma cells, suggesting that vector integration was required for high transduction. In conclusion, HIV vectors are promising tools for studies of gene functions in normal human B cells and myeloma cells for the purpose of research and the development of gene therapies.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
C. Frecha, C. Costa, C. Levy, D. Negre, S. J. Russell, A. Maisner, G. Salles, K.-W. Peng, F.-L. Cosset, and E. Verhoeyen
Efficient and stable transduction of resting B lymphocytes and primary chronic lymphocyte leukemia cells using measles virus gp displaying lentiviral vectors
Blood,
October 8, 2009;
114(15):
3173 - 3180.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Gimenez-Cassina, F. Lim, T. Cerrato, G. M. Palomo, and J. Diaz-Nido
Mitochondrial Hexokinase II Promotes Neuronal Survival and Acts Downstream of Glycogen Synthase Kinase-3
J. Biol. Chem.,
January 30, 2009;
284(5):
3001 - 3011.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. L. Good and S. G. Tangye
Decreased expression of Kruppel-like factors in memory B cells induces the rapid response typical of secondary antibody responses
PNAS,
August 14, 2007;
104(33):
13420 - 13425.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. N. Levasseur, T. M. Ryan, K. M. Pawlik, and T. M. Townes
Correction of a mouse model of sickle cell disease: lentiviral/antisickling {beta}-globin gene transduction of unmobilized, purified hematopoietic stem cells
Blood,
December 15, 2003;
102(13):
4312 - 4319.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
