000053774 001__ 53774
000053774 005__ 20190604054650.0
000053774 0247_ $$2doi$$a10.1016/j.ymthe.2005.05.010
000053774 037__ $$aARTICLE
000053774 245__ $$aTransduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors
000053774 269__ $$a2005
000053774 260__ $$c2005
000053774 336__ $$aJournal Articles
000053774 520__ $$aRecently, 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.
000053774 700__ $$aKvell, Krisztian
000053774 700__ $$aNguyen, Tuan H
000053774 700__ $$aSalmon, Patrick
000053774 700__ $$aGlauser, Frédéric
000053774 700__ $$aWerner-Favre, Christiane
000053774 700__ $$aBarnet, Marc
000053774 700__ $$aSchneider, Pascal
000053774 700__ $$0240083$$g167919$$aTrono, Didier
000053774 700__ $$aZubler, Rudolf H
000053774 773__ $$j12$$tMolecular Therapy$$q892-899
000053774 909C0 $$xU11172$$0252036$$pLVG
000053774 909CO $$pSV$$particle$$ooai:infoscience.tind.io:53774
000053774 917Z8 $$x148230
000053774 937__ $$aLVG-ARTICLE-2005-008
000053774 970__ $$a16005685/LVG
000053774 973__ $$rREVIEWED$$sPUBLISHED$$aEPFL
000053774 980__ $$aARTICLE