Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors

Krisztian Kvell, Tuan H. Nguyen, Patrick Salmon, Frédéric Glauser, Christiane Werner-Favre, Marc Barnet, Pascal Schneider, Didier Trono, Rudolf H. Zubler

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Recently, using HIV-1-derived lentivectors, we obtained efficient transduction of primary human B lymphocytes cocultured with murine EL-4 B5 thymoma cells, but not of isolated B cells activated by CD40 ligation. Coculture with a cell line is problematic for gene therapy applications or study of gene functions. We have now found that transduction of B cells in a system using CpG DNA was comparable to that in the EL-4 B5 system. A monocistronic vector with a CMV promoter gave 32 ± 4.7% green fluorescent protein (GFP)+ cells. A bicistronic vector, encoding IL-4 and GFP in the first and second cistrons, respectively, gave 14.2 ± 2.1% GFP+ cells and IL-4 secretion of 1.3 ± 0.2 ng/105 B cells/24 h. This was similar to results obtained in CD34+ cells using the elongation factor-1α promoter. Activated memory and naive B cells were transducible. After transduction with a bicistronic vector encoding a viral FLIP molecule, vFLIP was detectable by FACS or Western blot in GFP+, but not in GFP-, B cells, and 57% of sorted GFP+ B cells were protected against Fas ligand-induced cell death. This system should be useful for gene function research in primary B cells and development of gene therapies.

Original languageEnglish
Pages (from-to)892-899
Number of pages8
JournalMolecular Therapy
Volume12
Issue number5
DOIs
Publication statusPublished - Nov 2005

Keywords

  • Bicistronic vectors
  • CpG DNA
  • HIV-1-derived lentivectors
  • Human primary B lymphocytes
  • Viral FLIP

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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  • Cite this

    Kvell, K., Nguyen, T. H., Salmon, P., Glauser, F., Werner-Favre, C., Barnet, M., Schneider, P., Trono, D., & Zubler, R. H. (2005). Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors. Molecular Therapy, 12(5), 892-899. https://doi.org/10.1016/j.ymthe.2005.05.010