title: Generation of a lentiviral vector producer cell clone for human Wiskott-Aldrich syndrome gene therapy.
abstract: We have developed a producer cell line that generates lentiviral vector particles of high titer. The vector encodes the Wiskott-Aldrich syndrome (WAS) protein. An insulator element has been added to the long terminal repeats of the integrated vector to limit proto-oncogene activation. The vector provides high-level, stable expression of WAS protein in transduced murine and human hematopoietic cells. We have also developed a monoclonal antibody specific for intracellular WAS protein. This antibody has been used to monitor expression in blood and bone marrow cells after transfer into lineage negative bone marrow cells from WAS mice and in a WAS negative human B-cell line. Persistent expression of the transgene has been observed in transduced murine cells 12-20 weeks following transplantation. The producer cell line and the specific monoclonal antibody will facilitate the development of a clinical protocol for gene transfer into WAS protein deficient stem cells.
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Wielgosz
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Yoon-Sang
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Gael G
authors.initials: GG
authors.lastname: Carney
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhan
authors.affiliation: Department of Infectious Diseases, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Muralidhar
authors.initials: M
authors.lastname: Reddivari
authors.affiliation: Department of Infectious Diseases, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Terry
authors.initials: T
authors.lastname: Coop
authors.affiliation: Department of Infectious Diseases, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Heath
authors.affiliation: Immunology Department, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Brown
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St. Jude Children's Research Hospital , Memphis, Tennessee, USA.
authors.forename: Arthur W
authors.initials: AW
authors.lastname: Nienhuis