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

Article Article scientifique Article Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors eng 2005-11 author Kvell Krisztian 134466 author Nguyen Tuan H. author Salmon Patrick 505385 Services communs de la faculté de médecine (SCMU) author Glauser Frédéric 105961 author Favre Christiane 26436 author Barnet Marc author Schneider Rémi Pascal 30562 author Trono Didier 116006 author Zubler Rudolf 24774 892-899 10.1016/j.ymthe.2005.05.010 16005685 1525-0016 https://linkinghub.elsevier.com/retrieve/pii/S1525001605002273 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. 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 Dewey 616 56 Département de médecine 221 false Molecular therapy 12 5 ab3cbdb6-6750-4ffb-b064-3ed02644edc0 Article (Published version) Public Article-scientifique-from-unpaywall.pdf 324077 application/pdf CC BY-NC-ND-4.0 ec97ef7cd7e6f1bdb24c8373639b3f3960a31fe74357bcb400c7bae49ae352cc openAccess Faculté de médecine / Section de médecine clinique / Département de médecine

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Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors	Molecular therapy			2005	107	132

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Favre, Christiane