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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)85-93
Number of pages9
JournalGene Therapy
Volume11
Issue number1
DOIs
Publication statusPublished - Jan 2004

Keywords

  • Acute lymphoblastic leukemia
  • CD40L
  • Dendritic cells
  • Lentivirus

ASJC Scopus subject areas

  • Genetics

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