Gene transduction in human monocyte-derived dendritic cells using lentiviral vectors

Abstract

Monocyte-derived dendritic cells (MDDCs) are widely used in the fi eld of human immunology. Although a variety of gene delivery procedures have been used in MDDC, it has remained dif fi cult to achieve robust gene transductions. In this chapter, we describe a procedure for high ef fi ciency 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. © Springer Science+Business Media, LLC 2013.