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Prepublished online as a Blood First Edition Paper on October 24, 2002; DOI 10.1182/blood-2001-12-0249.
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Blood, 1 March 2003, Vol. 101, No. 5, pp. 1727-1733
GENE THERAPY
Efficient transduction of primary human B lymphocytes and
nondividing 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-François Arrighi,
Vincent Piguet,
Didier Trono, and
Rudolf H. Zubler
From the Division of Hematology, Department of
Medicine, Department of Genetics and Microbiology, and Department of
Dermatology, University Hospital, Geneva, Switzerland.
We studied the transduction of primary human B lymphocytes and
myeloma cells with lentiviral vectors. In peripheral blood B cells that
had been activated with helper T cells (murine thymoma EL-4 B5) and
cytokines, multiply attenuated HIV-1-derived vectors pseudotyped with
vesicular stomatitis virus (VSV) G-envelope protein achieved the
expression of green fluorescence 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 little variation. The human cytomegalovirus (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 primary 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 different results. Freshly isolated,
nondividing myeloma cells were efficiently transduced by HIV vectors;
for 6 myelomas the range was 14% to 77% (median, 28%)
GFP+ cells. HIV vectors with a mutant integrase led to no
significant GFP signal in primary 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
primary human B cells and myeloma cells for the purposes of research
and the development of gene therapies.
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