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

Fabrice Bovia, Patrick Salmon, Thomas Matthes, K. Kvell, Tuan H. Nguyen, Christiane Werner-Favre, Marc Barnet, Monika Nagy, Florence Leuba, Jean François Arrighi, Vincent Piguet, Didier Trono, Rudolf H. Zubler

Research output: Contribution to journalArticle

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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

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Lymphocytes
HIV-1
B-Lymphocytes
Cells
Fluorescence
HIV
Proteins
Viruses
Cytokines
Peptide Elongation Factor 1
Viral Envelope Proteins
Integrases
Gene therapy
CD40 Ligand
Thymoma
T-cells
Helper-Inducer T-Lymphocytes
Cytomegalovirus
Human Herpesvirus 4
Cell culture

ASJC Scopus subject areas

  • Hematology

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Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors. / Bovia, Fabrice; Salmon, Patrick; Matthes, Thomas; Kvell, K.; Nguyen, Tuan H.; Werner-Favre, Christiane; Barnet, Marc; Nagy, Monika; Leuba, Florence; Arrighi, Jean François; Piguet, Vincent; Trono, Didier; Zubler, Rudolf H.

In: Blood, Vol. 101, No. 5, 01.03.2003, p. 1727-1733.

Research output: Contribution to journalArticle

Bovia, F, Salmon, P, Matthes, T, Kvell, K, Nguyen, TH, Werner-Favre, C, Barnet, M, Nagy, M, Leuba, F, Arrighi, JF, Piguet, V, Trono, D & Zubler, RH 2003, 'Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors', Blood, vol. 101, no. 5, pp. 1727-1733. https://doi.org/10.1182/blood-2001-12-0249
Bovia, Fabrice ; Salmon, Patrick ; Matthes, Thomas ; Kvell, K. ; Nguyen, Tuan H. ; Werner-Favre, Christiane ; Barnet, Marc ; Nagy, Monika ; Leuba, Florence ; Arrighi, Jean François ; Piguet, Vincent ; Trono, Didier ; Zubler, Rudolf H. / Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors. In: Blood. 2003 ; Vol. 101, No. 5. pp. 1727-1733.
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