Efficient transduction of primary human B lymphocytes and nondividing myeloma B cells with HIV-1-derived lentiviral vectors
| Autor: | Didier Trono, Fabrice Bovia, Thomas Matthes, Vincent Piguet, Patrick Salmon, Jean-François Arrighi, Christiane Werner-Favre, Tuan H. Nguyen, Marc Barnet, Monika Nagy, Rudolf H. Zubler, Krisztian Kvell, Florence Leuba |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Herpesvirus 4
Human Genetic enhancement Recombinant Fusion Proteins Immunology CD40 Ligand Genetic Vectors Green Fluorescent Proteins Cytomegalovirus HIV Integrase Biology Lymphocyte Activation Biochemistry Virus Vesicular stomatitis Indiana virus Viral vector Green fluorescent protein Transduction (genetics) Peptide Elongation Factor 1 Genes Reporter Tumor Cells Cultured Humans Promoter Regions Genetic Cells Cultured B-Lymphocytes CD40 Genetic transfer Defective Viruses Cell Biology Hematology T-Lymphocytes Helper-Inducer biology.organism_classification Virology Genes gag Genes pol Genes rev Leukemia Virus Murine Luminescent Proteins Vesicular stomatitis virus Genes tat biology.protein HIV-1 Neoplastic Stem Cells Cytokines Multiple Myeloma |
| Zdroj: | Blood. 101(5) |
| ISSN: | 0006-4971 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|