Systematic improvements in lentiviral transduction of primary human natural killer cells undergoing ex vivo expansion
David Allan, Mala Chakraborty, Giacomo Waller, Michael J. Hochman, Akkapon Poolcharoen, Robert Reger, Richard Childs
Abstract
NK cells) and successful subsequent proliferation of the transduced cells. Investigation of multiple internal promoter sequences within the same lentiviral vector revealed differences in percentage and level of transgene expression per NK cell. Bicistronic lentiviral vectors encoding both GFP and proteins suitable for the isolation of transduced cells with magnetic beads led to efficient transgene expression in NK cells. The optimized approaches described herein provide a template for protocols that generate large numbers of fully functional and highly purified lentivirus-transduced NK cells for clinical trials.
Topics & Concepts
Transduction (biophysics)Vesicular stomatitis virusBiologyJanus kinase 3TransgeneViral vectorCell biologyGenetic enhancementEx vivoInterleukin 21Molecular biologyVirologyImmunologyT cellVirusIn vitroImmune systemRecombinant DNAGeneGeneticsBiochemistryImmune Cell Function and InteractionVirus-based gene therapy researchCAR-T cell therapy research