Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1–derived lentiviral vectors

F Bovia, P Salmon, T Matthes, K Kvell… - Blood, The Journal …, 2003 - ashpublications.org
F Bovia, P Salmon, T Matthes, K Kvell, TH Nguyen, C Werner-Favre, M Barnet, M Nagy…
Blood, The Journal of the American Society of Hematology, 2003ashpublications.org
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 …
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-1α 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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