title: Retrovirus-mediated WASP gene transfer corrects Wiskott-Aldrich syndrome T-cell dysfunction.
abstract: The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by thrombocytopenia, eczema, and immunodeficiency. At present, the only definitive therapy for the disease is allogeneic bone marrow transplantation (BMT). Because of the frequent lack of suitable donors and the potential severe complications associated with BMT, the development of gene-based therapeutic strategies for WAS is highly desirable. To study whether corrective gene transfer into WAS T cells can lead to restoration of the immunologic defects of WAS, a retroviral vector expressing the WAS protein (WASP) gene was used to transduce human T-lymphotropic virus type 1-transformed T-cell lines and primary T lymphocytes from patients with WAS. After transduction, WAS T cells showed levels of WASP expression similar to those found in cells from normal individuals. In addition, the reconstituted WASP interacted in vitro with proteins containing SH3 domain such as Grb2, PLC-gamma1, and Fyn, each of which are connected to signaling pathways linked to the actin cytoskeleton. Furthermore, after CD3 cross-linking, transduced WAS T lines showed improvement of actin polymerization and T-cell receptor/CD3 down-regulation. More importantly, primary WAS T lymphocytes transduced with WASP acquired the ability to proliferate in response to anti-CD3 stimulation. These findings suggest that biologic defects of WAS T cells can be corrected in vitro by retrovirus-mediated gene transfer and pose the basis for future investigation of gene therapy as treatment for WAS.
authors.affiliation: Disorders of Immunity Section, Genetics and Molecular Biology Branch, National Human Genome Research Institute,10 Center Drive, Building 10, Room 10C103, Bethesda, MD 20892-1851, USA.
authors.forename: Taizo
authors.initials: T
authors.lastname: Wada
authors.forename: G Jayashree
authors.initials: GJ
authors.lastname: Jagadeesh
authors.forename: David L
authors.initials: DL
authors.lastname: Nelson
authors.forename: Fabio
authors.initials: F
authors.lastname: Candotti
mesh_terms: D000199:Actins / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D002461:Cell Line, Transformed; D015536:Down-Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015368:Human T-lymphotropic virus 1 / Q000378:metabolism; D006801:Humans; D009154:Mutation; D011506:Proteins / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D014923:Wiskott-Aldrich Syndrome / Q000235:genetics* / Q000276:immunology / Q000628:therapy; D051316:Wiskott-Aldrich Syndrome Protein; D018909:src Homology Domains
chemical_list: D000199:Actins; D017252:CD3 Complex; D011506:Proteins; D011948:Receptors, Antigen, T-Cell; C495767:WAS protein, human; D051316:Wiskott-Aldrich Syndrome Protein