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 (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.
Bovia
Fabrice
F
Division of Hematology, Department of Medicine, University Hospital, Geneva, Switzerland.
Salmon
Patrick
P
Matthes
Thomas
T
Kvell
Krisztian
K
Nguyen
Tuan H
TH
Werner-Favre
Christiane
C
Barnet
Marc
M
Nagy
Monika
M
Leuba
Florence
F
Arrighi
Jean-Francois
JF
Piguet
Vincent
V
Trono
Didier
D
Zubler
Rudolf H
RH
D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D023201:CD40 Ligand / Q000494:pharmacology; D002478:Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000821:virology; D016207:Cytokines / Q000494:pharmacology; D003587:Cytomegalovirus / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics / Q000502:physiology*; D017930:Genes, Reporter; D015750:Genes, gag; D015751:Genes, pol; D015757:Genes, rev; D015756:Genes, tat; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D049452:Green Fluorescent Proteins; D019427:HIV Integrase / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D004854:Herpesvirus 4, Human / Q000502:physiology; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000502:physiology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation; D009101:Multiple Myeloma / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000821:virology*; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000821:virology; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics
D016207:Cytokines; D008164:Luminescent Proteins; D020648:Peptide Elongation Factor 1; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand; D049452:Green Fluorescent Proteins; D019427:HIV Integrase