Functional transfer of CD40L gene in human B-cell precursor ALL blasts by second-generation SIN lentivectors View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2003-12-18

AUTHORS

M Bonamino, M Serafini, G D'Amico, G Gaipa, E Todisco, S Bernasconi, J Golay, A Biondi, M Introna

ABSTRACT

Three different second-generation lentiviral self-inactivating vectors containing CMV, EF1α 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. More... »

PAGES

85-93

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/sj.gt.3302141

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