title: A simple, versatile and efficient method to genetically modify human monocyte-derived dendritic cells with HIV-1-derived lentiviral vectors.
abstract: Lentiviral vectors derived from the human immunodeficiency type 1 virus (HIV-1 LV) are among the finest tools available today for the genetic modification of human monocyte-derived dendritic cells (MDDCs). However, this process is largely inefficient because MDDCs show a strong resistance to HIV-1 transduction. Here we describe a step-by-step protocol from the production of LVs to cell transduction that allows the efficient genetic modification of MDDCs. This protocol can be completed in 23 d from the initial phase of LV production to the final analysis of the results of MDDC transduction. The method relies on the simultaneous addition of HIV-1 LVs along with noninfectious virion-like particles carrying Vpx, a nonstructural protein encoded by the simian immunodeficiency virus (Vpx-VLPs). When thus provided in target cells, Vpx exerts a strong positive effect on incoming LVs by counteracting the restriction present in MDDCs; accordingly, 100% of cells can be transduced with low viral inputs. Vpx-VLPs will improve the efficiency of LV-mediated transduction of MDDCs with vectors for both ectopic gene expression and depletion studies.
authors.affiliation: Department of Human Virology, Ecole Normale Supérieure de Lyon, France.
authors.forename: Grégory
authors.initials: G
authors.lastname: Berger
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Durand
authors.forename: Caroline
authors.initials: C
authors.lastname: Goujon
authors.forename: Xuan-Nhi
authors.initials: XN
authors.lastname: Nguyen
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Cordeil
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Darlix
authors.forename: Andrea
authors.initials: A
authors.lastname: Cimarelli
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology / Q000821:virology; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009000:Monocytes / Q000166:cytology; D014161:Transduction, Genetic / Q000379:methods*; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics; D014771:Virion / Q000235:genetics
chemical_list: C513059:VPX protein, Simian immunodeficiency virus; D054334:Viral Regulatory and Accessory Proteins