Repopulation of B-lymphocytes with restricted gene expression using haematopoietic stem cells engineered with lentiviral vectors
| Autor: | Rewas Fatah, Taher E. Taher, Rizgar A. Mageed, David Gould, Fulvio D'Acquisto, C Tulone |
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| Rok vydání: | 2008 |
| Předmět: |
Transgene
Genetic enhancement Antigens CD19 Genetic Vectors Green Fluorescent Proteins Gene Expression Mice SCID Biology CD19 Article Viral vector Cell Line Mice Transduction Genetic Gene expression Genetics Animals Humans Transgenes Enhancer Promoter Regions Genetic Molecular Biology B-Lymphocytes Mice Inbred BALB C Lentivirus Hematopoietic Stem Cell Transplantation Genetic Therapy Flow Cytometry Hematopoietic Stem Cells Molecular biology Haematopoiesis biology.protein Molecular Medicine Stem cell Genetic Engineering |
| Zdroj: | Gene therapy. 15(13) |
| ISSN: | 1476-5462 |
| Popis: | B-lymphocytes play a key role in the pathogenesis of many immune-mediated diseases, such as autoimmune and atopic diseases. Therefore, targeting B-lymphocytes provides a rationale for refining strategies to treat such diseases for long-term clinical benefits and minimal side effects. In this study we describe a protocol for repopulating irradiated mice with B-lymphocytes engineered for restricted expression of transgenes using haematopoietic stem cells. A self-inactivating lentiviral vector, which encodes enhanced green fluorescence protein (EGFP) from the spleen focus-forming virus (SFFV) promoter, was used to generate new vectors that permit restricted EGFP expression in B-lymphocytes. To achieve this, the SFFV promoter was replaced with the B-lymphocyte-restricted CD19 promoter. Further, an immunoglobulin heavy chain enhancer (Emu) flanked by the associated matrix attachment regions (MARs) was inserted upstream of the CD19 promoter. Incorporation of the Emu-MAR elements upstream of the CD19 promoter resulted in enhanced, stable and selective transgene expression in human and murine B-cell lines. In addition, this modification permitted enhanced selective EGFP expression in B-lymphocytes in vivo in irradiated mice repopulated with transduced bone marrow haematopoietic stem cells (BMHSCs). The study provides evidence for the feasibility of targeting B-lymphocytes for therapeutic restoration of normal B-lymphocyte functions in patients with B-cell-related diseases. |
| Databáze: | OpenAIRE |
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