title: Maximal lentivirus-mediated gene transfer and sustained transgene expression in human hematopoietic primitive cells and their progeny.
abstract: Gene therapy using permanent modifications of hematopoietic stem cells (HSC) has increasing potential applications for both genetic and acquired diseases. Considerable progress has been made recently in gene transfer to HSC by the use of lentivirus-derived vectors, which have the capacity to transduce noncycling cells. However, overall efficiency of HSC transduction reported so far is still not sufficient for numerous applications. We describe here an improved HSC transduction protocol, using the previously described lentiviral vector, that leads to more than 90% transduction of human CD34+ cells from cord blood, including NOD-LtSz-scid/scid repopulating cells. Moreover, under the same conditions, we transduce more than 75% and 80% of CD34+ cells mobilized in peripheral blood and from bone marrow, respectively. We further show that transgene expression is stable through time and hematopoietic cell differentiation in vitro as well as in vivo. Such a high HSC transduction efficiency opens new opportunities for both gene therapy applications and functional studies of regulator proteins of hematopoiesis.
authors.affiliation: Institut Cochin, INSERM U567, CNRS UMR 8104, Hematology Department, Maternité de Port-Royal, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Amsellem
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Ravet
authors.forename: Serge
authors.initials: S
authors.lastname: Fichelson
authors.forename: Françoise
authors.initials: F
authors.lastname: Pflumio
authors.forename: Anne
authors.initials: A
authors.lastname: Dubart-Kupperschmitt
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis / Q000235:genetics; D001854:Bone Marrow Cells / Q000378:metabolism; D002469:Cell Separation; D005312:Fetal Blood / Q000166:cytology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D016133:Polymerase Chain Reaction; D019076:Transgenes
chemical_list: D018952:Antigens, CD34; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins