Document Detail

Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors.
MedLine Citation:
PMID:  16005685     Owner:  NLM     Status:  MEDLINE    
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.
Krisztian Kvell; Tuan H Nguyen; Patrick Salmon; Frédéric Glauser; Christiane Werner-Favre; Marc Barnet; Pascal Schneider; Didier Trono; Rudolf H Zubler
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-07-11
Journal Detail:
Title:  Molecular therapy : the journal of the American Society of Gene Therapy     Volume:  12     ISSN:  1525-0016     ISO Abbreviation:  Mol. Ther.     Publication Date:  2005 Nov 
Date Detail:
Created Date:  2005-10-31     Completed Date:  2006-01-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  100890581     Medline TA:  Mol Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  892-9     Citation Subset:  IM    
Division of Hematology, Department of Internal Medicine, University Hospitals, 1211 Geneva-14, Switzerland.
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MeSH Terms
Antigens, CD34
B-Lymphocytes / immunology,  metabolism*
CD40 Ligand
Cell Culture Techniques
Cells, Cultured
CpG Islands*
Genetic Vectors*
Green Fluorescent Proteins / genetics,  metabolism
HIV-1 / genetics*
Interleukin-4 / secretion*
Lentivirus / genetics*
Molluscum contagiosum virus / genetics
Multiple Myeloma
Transduction, Genetic*
Reg. No./Substance:
0/Antigens, CD34; 147205-72-9/CD40 Ligand; 147336-22-9/Green Fluorescent Proteins; 207137-56-2/Interleukin-4

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