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

K. 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

Fingerprint

B-Lymphocytes
Green Fluorescent Proteins
DNA
Interleukin-4
Genetic Therapy
Genes
Peptide Elongation Factor 1
Fas Ligand Protein
Thymoma
Coculture Techniques
Ligation
HIV-1
Cell Death
Western Blotting
Cell Line
Research

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Kvell, K., Nguyen, T. H., Salmon, P., Glauser, F., Werner-Favre, C., Barnet, M., ... 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

Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors. / Kvell, K.; Nguyen, Tuan H.; Salmon, Patrick; Glauser, Frédéric; Werner-Favre, Christiane; Barnet, Marc; Schneider, Pascal; Trono, Didier; Zubler, Rudolf H.

In: Molecular Therapy, Vol. 12, No. 5, 11.2005, p. 892-899.

Research output: Contribution to journalArticle

Kvell, K, Nguyen, TH, Salmon, P, Glauser, F, Werner-Favre, C, Barnet, M, Schneider, P, Trono, D & Zubler, RH 2005, 'Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors', Molecular Therapy, vol. 12, no. 5, pp. 892-899. https://doi.org/10.1016/j.ymthe.2005.05.010
Kvell, K. ; Nguyen, Tuan H. ; Salmon, Patrick ; Glauser, Frédéric ; Werner-Favre, Christiane ; Barnet, Marc ; Schneider, Pascal ; Trono, Didier ; Zubler, Rudolf H. / Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors. In: Molecular Therapy. 2005 ; Vol. 12, No. 5. pp. 892-899.
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