title: Gene transduction in human monocyte-derived dendritic cells using lentiviral vectors.
abstract: Monocyte-derived dendritic cells (MDDCs) are widely used in the field of human immunology. Although a variety of gene delivery procedures have been used in MDDC, it has remained difficult to achieve robust gene transductions. In this chapter, we describe a procedure for high efficiency gene transduction in human MDDCs using lentiviral vectors. Gene transduction based on HIV-1-derived lentiviral vectors is restricted at the level of reverse transcription by the cellular protein SAMHD1 in MDDCs. Co-transduction of the MDDCs with helper particles derived from SIVmac that contain the viral protein Vpx removes this restriction, leading to a drastic increase in the rate of gene transduction. This procedure leads to nontoxic, efficient and stable transduction in MDDCs. It can be applied to any HIV-1-derived lentiviral vector, including shRNA lentiviral vectors for RNAi. Transduced MDDCs are not activated by the transduction and can be activated normally by TLR ligands.
authors: Institut Curie, INSERM U932, Paris, France.
authors: Takeshi
authors: T
authors: Satoh
authors: Institut Curie, INSERM U932, Paris, France. nicolas.manel@curie.fr.
authors: Nicolas
authors: N
authors: Manel
mesh_terms: D002454:Cell Differentiation; D002469:Cell Separation; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009000:Monocytes / Q000166:cytology*; D015302:Simian Immunodeficiency Virus / Q000502:physiology; D014161:Transduction, Genetic / Q000379:methods*