High levels of transduction of human dendritic cells with optimized SIV vectors.
As major antigen-presenting cells and effectors in the maintenance of tolerance, dendritic cells (DCs) are key cells of the immune system and can thus be envisioned to have roles in immunotherapy strategies. We, and others, previously showed that simian immunodeficiency virus (SIV)-derived lentiviral vectors were able to deliver a gene into human differentiated DCs. We describe here the upgrading of the SIV vector system and the improvements of the transduction protocol, which allowed us to transduce more than 90% of human monocyte-derived DCs. We developed new SIV lentiviral vectors carrying SIV splice regulatory elements and either the woodchuck hepatitis virus regulatory element (WPRE) or the murine phosphoglycerate-kinase 1 (PGK) promoter. We show that insertion of the WPRE in the SIV vector is detrimental to gene transfer in DCs, while this sequence increases transgene expression in 293T cells. Using an optimized SIV vector, high levels of transgene expression were obtained in more than 30% of human DCs at a multiplicity of infection (MOI) of 1, and close to 100% using a MOI of 20. VSV-G pseudotyped vectors generated with only gag, pol, tat, and rev helper functions failed to transduce DCs. This defect was completely rescued when the SIV accessory gene vpx was provided in trans in vector-producing cells. Genetically modified DCs were shown to behave as bona fide DCs in both allogenic and autologous mixed leukocyte reactions. These findings allow us to propose an optimal system for efficient and safe DC transduction with improved SIV vectors.
Mangeot
Philippe-Emmanuel
PE
Unité de Virologie Humaine Inserm U412, Labo Rétro, ENS Lyon, France.
Duperrier
Karine
K
Nègre
Didier
D
Boson
Bertrand
B
Rigal
Dominique
D
Cosset
François-Loïc
FL
Darlix
Jean-Luc
JL
D003713:Dendritic Cells / Q000502:physiology*; D005822:Genetic Vectors; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006801:Humans; D010735:Phosphoglycerate Kinase / Q000235:genetics; D011401:Promoter Regions, Genetic; D015302:Simian Immunodeficiency Virus; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000235:genetics
D014764:Viral Proteins; D010735:Phosphoglycerate Kinase