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

Article Article scientifique Article Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors eng 2002-10-24 2003-03-01 author Bovia Fabrice 29526 author Salmon Patrick 505385 author Matthes Thomas 27368 author Kvell Krisztian 134466 author Nguyen Tuan Huy 133497 author Werner-Favre Christiane author Barnet Marc author Nagy Monika 514344 author Leuba Florence 96535 author Arrighi Jean-François 28404 author Piguet Vincent 84926 author Trono Didier 116006 author Zubler Rudolf H 1727-1733 10.1182/blood-2001-12-0249 12406892 0006-4971 <p>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-1alpha 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.</p> B-Lymphocytes / drug effects B-Lymphocytes / immunology B-Lymphocytes / metabolism B-Lymphocytes / virology CD40 Ligand / pharmacology Cells, Cultured / drug effects Cells, Cultured / immunology Cells, Cultured / metabolism Cells, Cultured / virology Cytokines / pharmacology Cytomegalovirus / genetics Defective Viruses / genetics Defective Viruses / physiology Genes, Reporter Genes, gag Genes, pol Genes, rev Genes, tat Genetic Vectors / genetics Genetic Vectors / physiology Green Fluorescent Proteins HIV Integrase / deficiency HIV Integrase / genetics HIV Integrase / physiology HIV-1 / genetics HIV-1 / physiology Herpesvirus 4, Human / physiology Humans Leukemia Virus, Murine / genetics Leukemia Virus, Murine / physiology Luminescent Proteins / biosynthesis Luminescent Proteins / genetics Lymphocyte Activation Multiple Myeloma / pathology Neoplastic Stem Cells / drug effects Neoplastic Stem Cells / metabolism Neoplastic Stem Cells / virology Peptide Elongation Factor 1 / genetics Peptide Elongation Factor 1 / physiology Promoter Regions, Genetic Recombinant Fusion Proteins / biosynthesis T-Lymphocytes, Helper-Inducer / immunology Tumor Cells, Cultured / drug effects Tumor Cells, Cultured / immunology Tumor Cells, Cultured / metabolism Tumor Cells, Cultured / virology Vesicular stomatitis Indiana virus / genetics Dewey 540 Dewey 616 56 Département de médecine 221 false 586 Analyses cellulaires et moléculaires des hémopathies malignes 929 231 TRONO DIDIER 296 Blood 101 5 b72c163c-dfc5-49bb-ad1f-f7183ebe1679 Article (Published version) Public 1-s2.0-S0006497120508628-main.pdf 244193 application/pdf OA OTHER openAccess Faculté de médecine / Section de médecine clinique / Département de médecine

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Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors	Molecular therapy	2005	107	132
Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors	Blood	2003	342	220

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