title: Characterization of novel safe lentiviral vectors derived from simian immunodeficiency virus (SIVmac251) that efficiently transduce mature human dendritic cells.
abstract: We describe the generation and the characterization of new lentiviral vectors derived from SIVmac251, a simian immunodeficiency virus (SIV). A methodical approach was used to engineer both efficient and safe packaging constructs allowing the production of SIV viral core proteins. SIV-vectors encoding GFP (green fluorescent protein) were generated as VSV-G-pseudotyped particles upon transient expression of the vector construct and helper functions in 293 cells. The SIV vectors were able to transduce efficiently various target cell types at low multiplicity of infection, including monocyte-differentiated human dendritic cells (DCs) which retained their capacity to differentiate into mature DCs after gene transfer. Transduction of the DCs by the SIV vectors was prevented when infections were performed in the presence of AZT, a reverse-transcriptase inhibitor. After gene transfer, expression of the GFP in the target cells remained constant after several weeks, indicating that the vectors had been stably integrated into the genome of the host cells. Preparations of SIV vectors were systematically checked for the absence of replication-competent and recombinant retroviruses but remained negative, suggesting the innocuousness of these novel gene delivery vectors. Side-to-side comparisons with vectors derived from HIV-1 (human immunodeficiency virus) indicated that the SIV vectors were equally potent in transducing proliferating target cells. Finally, we have determined the infectivity of SIV vectors pseudotyped with surface glycoproteins of several membrane-enveloped viruses.
authors.affiliation: Vectorologie Rétrovirale and Thérapie Génique, U412 INSERM, IFR 74 and ENS de Lyon, France.
authors.forename: D
authors.initials: D
authors.lastname: Nègre
authors.forename: P E
authors.initials: PE
authors.lastname: Mangeot
authors.forename: G
authors.initials: G
authors.lastname: Duisit
authors.forename: S
authors.initials: S
authors.lastname: Blanchard
authors.forename: P O
authors.initials: PO
authors.lastname: Vidalain
authors.forename: P
authors.initials: P
authors.lastname: Leissner
authors.forename: A J
authors.initials: AJ
authors.lastname: Winter
authors.forename: C
authors.initials: C
authors.lastname: Rabourdin-Combe
authors.forename: M
authors.initials: M
authors.lastname: Mehtali
authors.forename: P
authors.initials: P
authors.lastname: Moullier
authors.forename: J L
authors.initials: JL
authors.lastname: Darlix
authors.forename: F L
authors.initials: FL
authors.lastname: Cosset
mesh_terms: D000818:Animals; D002460:Cell Line; D003713:Dendritic Cells / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D015302:Simian Immunodeficiency Virus / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D022701:Virosomes
chemical_list: D008164:Luminescent Proteins; D022701:Virosomes; D049452:Green Fluorescent Proteins