title: Expression of human glucocerebrosidase following retroviral vector-mediated transduction of murine hematopoietic stem cells.
abstract: The human glucocerebrosidase (GC) gene has been expressed in the progeny of murine hematopoietic stem cells following transduction of marrow with a retroviral vector (G2) containing the human GC cDNA. Murine marrow was transduced via co-cultivation following prestimulation in the presence or absence of recombinant IL-3 and IL-6. A high rate of gene transfer and expression (95%) was demonstrated in primary day 12 CFU-S foci following bone marrow transplantation (BMT) of G2-transduced marrow into lethally irradiated syngeneic recipient mice. Immunoreactive human GC protein was also documented in the CFU-S foci. Primary recipient mice were examined 4-6 months following BMT. A higher rate of gene transfer (87%) was seen in hematopoietic organs of recipients of prestimulated donor marrow compared with organs from initially unstimulated marrow (25%). A high rate of expression of human GC was also documented in the prestimulated organs (50%) when compared with the unstimulated group (25%). Secondary BMT was performed using marrow from the long-lived primary recipients. The human GC gene was present in 88% of secondary day 12 CFU-S foci examined in the prestimulated group versus 23% in the unstimulated group. Expression of the human GC gene was documented in secondary day 12 CFU-S foci, providing strong evidence of initial hematopoietic stem cell transduction.
authors.affiliation: Division of Research Immunology/BMT, University of Southern California School of Medicine, Children's Hospital, Los Angeles 90027.
authors.forename: J
authors.initials: J
authors.lastname: Weinthal
authors.forename: J A
authors.initials: JA
authors.lastname: Nolta
authors.forename: X J
authors.initials: XJ
authors.lastname: Yu
authors.forename: J
authors.initials: J
authors.lastname: Lilley
authors.forename: L
authors.initials: L
authors.lastname: Uribe
authors.forename: D B
authors.initials: DB
authors.lastname: Kohn
mesh_terms: D000818:Animals; D001483:Base Sequence; D016026:Bone Marrow Transplantation; D004247:DNA / Q000235:genetics; D005260:Female; D005776:Gaucher Disease / Q000628:therapy; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D005962:Glucosylceramidase / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000201:enzymology*; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D015345:Oligonucleotide Probes; D004247:DNA; D005962:Glucosylceramidase