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

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...

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Veröffentlicht in:Blood 2003-03, Vol.101 (5), p.1727-1733
Hauptverfasser: Bovia, Fabrice, Salmon, Patrick, Matthes, Thomas, Kvell, Krisztian, Nguyen, Tuan H., Werner-Favre, Christiane, Barnet, Marc, Nagy, Monika, Leuba, Florence, Arrighi, Jean-François, Piguet, Vincent, Trono, Didier, Zubler, Rudolf H.
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container_end_page 1733
container_issue 5
container_start_page 1727
container_title Blood
container_volume 101
creator Bovia, Fabrice
Salmon, Patrick
Matthes, Thomas
Kvell, Krisztian
Nguyen, Tuan H.
Werner-Favre, Christiane
Barnet, Marc
Nagy, Monika
Leuba, Florence
Arrighi, Jean-François
Piguet, Vincent
Trono, Didier
Zubler, Rudolf H.
description 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.
doi_str_mv 10.1182/blood-2001-12-0249
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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.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood-2001-12-0249</identifier><identifier>PMID: 12406892</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy ; Applied cell therapy and gene therapy ; B-Lymphocytes - drug effects ; B-Lymphocytes - immunology ; B-Lymphocytes - metabolism ; B-Lymphocytes - virology ; Biological and medical sciences ; Biotechnology ; 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 ; Fundamental and applied biological sciences. Psychology ; Gene therapy ; Genes, gag ; Genes, pol ; Genes, Reporter ; Genes, rev ; Genes, tat ; Genetic Vectors - genetics ; Genetic Vectors - physiology ; Green Fluorescent Proteins ; Health. Pharmaceutical industry ; Herpesvirus 4, Human - physiology ; HIV Integrase - deficiency ; HIV Integrase - genetics ; HIV Integrase - physiology ; HIV-1 - genetics ; HIV-1 - physiology ; Humans ; Industrial applications and implications. Economical aspects ; Leukemia Virus, Murine - genetics ; Leukemia Virus, Murine - physiology ; Luminescent Proteins - biosynthesis ; Luminescent Proteins - genetics ; Lymphocyte Activation ; Medical sciences ; 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 ; Transfusions. Complications. Transfusion reactions. 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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.</description><subject>Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy</subject><subject>Applied cell therapy and gene therapy</subject><subject>B-Lymphocytes - drug effects</subject><subject>B-Lymphocytes - immunology</subject><subject>B-Lymphocytes - metabolism</subject><subject>B-Lymphocytes - virology</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>CD40 Ligand - pharmacology</subject><subject>Cells, Cultured - drug effects</subject><subject>Cells, Cultured - immunology</subject><subject>Cells, Cultured - metabolism</subject><subject>Cells, Cultured - virology</subject><subject>Cytokines - pharmacology</subject><subject>Cytomegalovirus - genetics</subject><subject>Defective Viruses - genetics</subject><subject>Defective Viruses - physiology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene therapy</subject><subject>Genes, gag</subject><subject>Genes, pol</subject><subject>Genes, Reporter</subject><subject>Genes, rev</subject><subject>Genes, tat</subject><subject>Genetic Vectors - genetics</subject><subject>Genetic Vectors - physiology</subject><subject>Green Fluorescent Proteins</subject><subject>Health. Pharmaceutical industry</subject><subject>Herpesvirus 4, Human - physiology</subject><subject>HIV Integrase - deficiency</subject><subject>HIV Integrase - genetics</subject><subject>HIV Integrase - physiology</subject><subject>HIV-1 - genetics</subject><subject>HIV-1 - physiology</subject><subject>Humans</subject><subject>Industrial applications and implications. 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Intensive care medicine. Transfusions. Cell therapy and gene therapy</topic><topic>Applied cell therapy and gene therapy</topic><topic>B-Lymphocytes - drug effects</topic><topic>B-Lymphocytes - immunology</topic><topic>B-Lymphocytes - metabolism</topic><topic>B-Lymphocytes - virology</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>CD40 Ligand - pharmacology</topic><topic>Cells, Cultured - drug effects</topic><topic>Cells, Cultured - immunology</topic><topic>Cells, Cultured - metabolism</topic><topic>Cells, Cultured - virology</topic><topic>Cytokines - pharmacology</topic><topic>Cytomegalovirus - genetics</topic><topic>Defective Viruses - genetics</topic><topic>Defective Viruses - physiology</topic><topic>Fundamental and applied biological sciences. 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Economical aspects</topic><topic>Leukemia Virus, Murine - genetics</topic><topic>Leukemia Virus, Murine - physiology</topic><topic>Luminescent Proteins - biosynthesis</topic><topic>Luminescent Proteins - genetics</topic><topic>Lymphocyte Activation</topic><topic>Medical sciences</topic><topic>Multiple Myeloma - pathology</topic><topic>Neoplastic Stem Cells - drug effects</topic><topic>Neoplastic Stem Cells - metabolism</topic><topic>Neoplastic Stem Cells - virology</topic><topic>Peptide Elongation Factor 1 - genetics</topic><topic>Peptide Elongation Factor 1 - physiology</topic><topic>Promoter Regions, Genetic</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>T-Lymphocytes, Helper-Inducer - immunology</topic><topic>Transfusions. Complications. Transfusion reactions. Cell and gene therapy</topic><topic>Tumor Cells, Cultured - drug effects</topic><topic>Tumor Cells, Cultured - immunology</topic><topic>Tumor Cells, Cultured - metabolism</topic><topic>Tumor Cells, Cultured - virology</topic><topic>Vesicular stomatitis Indiana virus - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bovia, Fabrice</creatorcontrib><creatorcontrib>Salmon, Patrick</creatorcontrib><creatorcontrib>Matthes, Thomas</creatorcontrib><creatorcontrib>Kvell, Krisztian</creatorcontrib><creatorcontrib>Nguyen, Tuan H.</creatorcontrib><creatorcontrib>Werner-Favre, Christiane</creatorcontrib><creatorcontrib>Barnet, Marc</creatorcontrib><creatorcontrib>Nagy, Monika</creatorcontrib><creatorcontrib>Leuba, Florence</creatorcontrib><creatorcontrib>Arrighi, Jean-François</creatorcontrib><creatorcontrib>Piguet, Vincent</creatorcontrib><creatorcontrib>Trono, Didier</creatorcontrib><creatorcontrib>Zubler, Rudolf H.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bovia, Fabrice</au><au>Salmon, Patrick</au><au>Matthes, Thomas</au><au>Kvell, Krisztian</au><au>Nguyen, Tuan H.</au><au>Werner-Favre, Christiane</au><au>Barnet, Marc</au><au>Nagy, Monika</au><au>Leuba, Florence</au><au>Arrighi, Jean-François</au><au>Piguet, Vincent</au><au>Trono, Didier</au><au>Zubler, Rudolf H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1–derived lentiviral vectors</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>2003-03-01</date><risdate>2003</risdate><volume>101</volume><issue>5</issue><spage>1727</spage><epage>1733</epage><pages>1727-1733</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><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.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>12406892</pmid><doi>10.1182/blood-2001-12-0249</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record>
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subjects Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Applied cell therapy and gene therapy
B-Lymphocytes - drug effects
B-Lymphocytes - immunology
B-Lymphocytes - metabolism
B-Lymphocytes - virology
Biological and medical sciences
Biotechnology
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
Fundamental and applied biological sciences. Psychology
Gene therapy
Genes, gag
Genes, pol
Genes, Reporter
Genes, rev
Genes, tat
Genetic Vectors - genetics
Genetic Vectors - physiology
Green Fluorescent Proteins
Health. Pharmaceutical industry
Herpesvirus 4, Human - physiology
HIV Integrase - deficiency
HIV Integrase - genetics
HIV Integrase - physiology
HIV-1 - genetics
HIV-1 - physiology
Humans
Industrial applications and implications. Economical aspects
Leukemia Virus, Murine - genetics
Leukemia Virus, Murine - physiology
Luminescent Proteins - biosynthesis
Luminescent Proteins - genetics
Lymphocyte Activation
Medical sciences
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
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
Tumor Cells, Cultured - drug effects
Tumor Cells, Cultured - immunology
Tumor Cells, Cultured - metabolism
Tumor Cells, Cultured - virology
Vesicular stomatitis Indiana virus - genetics
title Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1–derived lentiviral vectors
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