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- Title
Functional transfer of CD40L gene in human B-cell precursor ALL blasts by second-generation SIN lentivectors.
- Authors
Bonamino, M.; Serafini, M.; D'Amico, G.; Gaipa, G.; Todisco, E.; Bernasconi, S.; Golay, J.; Biondi, A.; Introna, M.
- Abstract
Three different second-generation lentiviral self-inactivating vectors containing CMV, EF1a and PGK promoter, respectively, and all carrying the exogenous GFP gene, were compared for expression in human B-cell precursor ALL blasts. At a comparable percentage of transduction and vector DNA copy number, CMV clearly showed better efficiency of transcription. Human bone marrow stromal cells were favored compared to the MRC-5 cell line, as support for cell viability during infection. Cells were infected and analyzed after variable culture times ranging from 4 to 12 days, to reduce the possibility of pseudotransduction. In 10/14 samples, we detected more than 20% GFP-positive cells after exposure to high-titer viral supernatants. We then tested a similar vector carrying the human CD40L cDNA and, in similar infection conditions, obtained more than 20% transduction in 6/6 samples. The levels of transduction obtained were sufficient to induce the upregulation of CD83 molecule in cocultured immature dendritic cells.Gene Therapy (2004) 11, 85-93. doi:10.1038/sj.gt.3302141
- Subjects
GENETIC vectors; LENTIVIRUSES; PROMOTERS (Genetics); LYMPHOBLASTIC leukemia; B cells; GENE expression
- Publication
Gene Therapy, 2004, Vol 11, Issue 1, p85
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/sj.gt.3302141