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Scientific article
Open access
English

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

Published inMolecular therapy, vol. 12, no. 5, p. 892-899
Publication date2005-11
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/10(5) B cells/24 h. This was similar to results obtained in CD34+ cells using the elongation factor-1alpha 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.

eng
Keywords
  • Antigens, CD34
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • CD40 Ligand
  • Cell Culture Techniques
  • Cells, Cultured
  • CpG Islands
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HIV-1 / genetics
  • Humans
  • Interleukin-4 / metabolism
  • Lentivirus / genetics
  • Molluscum contagiosum virus / genetics
  • Multiple Myeloma
  • Transduction, Genetic
Citation (ISO format)
KVELL, Krisztian et al. Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors. In: Molecular therapy, 2005, vol. 12, n° 5, p. 892–899. doi: 10.1016/j.ymthe.2005.05.010
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Article (Published version)
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ISSN of the journal1525-0016
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