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- Title
Repopulation of B-lymphocytes with restricted gene expression using haematopoietic stem cells engineered with lentiviral vectors.
- Authors
Taher, T. E.; Tulone, C.; Fatah, R.; D'Acquisto, F.; Gould, D. J.; Mageed, R. A.
- Abstract
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 (Eμ) flanked by the associated matrix attachment regions (MARs) was inserted upstream of the CD19 promoter. Incorporation of the Eμ-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.Gene Therapy (2008) 15, 998–1006; doi:10.1038/gt.2008.33; published online 20 March 2008
- Subjects
GENE therapy; GENETIC engineering; THERAPEUTICS; LYMPHOCYTES; BLOOD cells; DISEASES; TRANSGENES; STEM cells; IMMUNOGLOBULIN genes
- Publication
Gene Therapy, 2008, Vol 15, Issue 13, p998
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/gt.2008.33