title: Comparison of various envelope proteins for their ability to pseudotype lentiviral vectors and transduce primitive hematopoietic cells from human blood.
abstract: Substantial effort has been invested in developing methodologies for efficient gene transfer into human, repopulating, hematopoietic stem cells. Oncoretroviral vectors are limited by the lack of nuclear mitosis in quiescent stem cells during ex vivo transduction, whereas the preintegration complex of lentiviral vectors contains nuclear-localizing signals that permit genome integration without mitosis. We have developed a flexible and versatile system for generating lentiviral vector particles and have pseudotyped such particles with amphotropic, ecotropic, feline endogenous virus (RD114) or vesicular stomatitis virus (VSV-G) envelope proteins. Particles of all four types could be concentrated approximately 100-fold by ultracentrifugation or ultrafiltration. RD114 or amphotropic particles were more efficient than VSV-G-pseudotyped particles at transducing human cord blood CD34(+) cells and clonogenic progenitors within that population. Amphotropic particles transduced cytokine-mobilized, human peripheral blood CD34(+) cells capable of establishing hematopoiesis in immunodeficient mice more efficiently than the other two types of particles. We conclude that the use of amphotropic pseudotyped lentiviral vector particles rather than the commonly used VSV-G-pseudotyped particles should be considered in potential applications of lentiviral vectors for gene transfer into this therapeutically relevant target cell population.
authors.affiliation: Department of Hematology/Oncology, Division of Experimental Hematology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
authors.forename: Hideki
authors.initials: H
authors.lastname: Hanawa
authors.forename: Patrick F
authors.initials: PF
authors.lastname: Kelly
authors.forename: Amit C
authors.initials: AC
authors.lastname: Nathwani
authors.forename: Derek A
authors.initials: DA
authors.lastname: Persons
authors.forename: Jody A
authors.initials: JA
authors.lastname: Vandergriff
authors.forename: Phillip
authors.initials: P
authors.lastname: Hargrove
authors.forename: Elio F
authors.initials: EF
authors.lastname: Vanin
authors.forename: Arthur W
authors.initials: AW
authors.lastname: Nienhuis
mesh_terms: D000818:Animals; D002415:Cats; D005822:Genetic Vectors / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D016086:Lentivirus; D051379:Mice; D016513:Mice, SCID; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins
chemical_list: D014759:Viral Envelope Proteins