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, Rudolf H. Zubler

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59 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1727-1733
Number of pages7
JournalBlood
Volume101
Issue number5
DOIs
Publication statusPublished - Mar 1 2003

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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    Bovia, F., Salmon, P., Matthes, T., Kvell, K., Nguyen, T. H., Werner-Favre, C., Barnet, M., Nagy, M., Leuba, F., Arrighi, J. F., Piguet, V., Trono, D., & Zubler, R. H. (2003). Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors. Blood, 101(5), 1727-1733. https://doi.org/10.1182/blood-2001-12-0249