title: CD34+ peripheral blood progenitors as a target for genetic correction of the two flavocytochrome b558 defective forms of chronic granulomatous disease.
abstract: Chronic granulomatous disease (CGD) can result from any of four single gene defects involving components of the superoxide (O2-.)-generating phagocyte NADPH oxidase (phox). The phox transmembrane flavocytochrome b558 is composed of two peptides, gp91phox and p22phox. Mutations of gp91phox cause X-linked CGD, whereas mutations of p22phox cause one of the three autosomal recessive forms of CGD. We used the Maloney leukemia virus-based MFG retrovirus vector to produce replication defective retroviruses encoding gp91phox or p22phox. To maximize viral titer MFG retroviruses do not contain internal promoter or resistance elements. Epstein-Barr virus transformed B-lymphocyte cell lines (EBV-B) derived from normal individuals contain phox components and produce O2-., whereas those derived from CGD patients show the CGD defect. Transduction of gp91phox or p22phox-deficient CGD EBV-B lines resulted in correction of O2-. production from a barely detectable baseline to an average 7.2% and 13.8% of normal control, respectively, without any selective regimen to enrich for transduced cells. CD34+ hematopoietic progenitor cells, the therapeutic target for gene therapy of CGD, were isolated from peripheral blood of CGD patients, transduced with MFG-phox retroviruses, and differentiated in culture to mature phagocytes. Transduction of progenitors corrected the gp91phox (seven patients) and p22phox (two patients) CGD phagocyte oxidase defect to 2.5% and 4.9% of normal O2-. production, respectively, representing an 87-fold and 161-fold increase. These studies show correction of flavocytochrome b558-deficient CGD in primary hematopoietic progenitors, providing a basis for development of gene therapy for the X-linked gp91phox and autosomal p22phox-deficient forms of CGD.
authors.affiliation: Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: F
authors.initials: F
authors.lastname: Li
authors.forename: G F
authors.initials: GF
authors.lastname: Linton
authors.forename: S
authors.initials: S
authors.lastname: Sekhsaria
authors.forename: N
authors.initials: N
authors.lastname: Whiting-Theobald
authors.forename: J P
authors.initials: JP
authors.lastname: Katkin
authors.forename: J I
authors.initials: JI
authors.lastname: Gallin
authors.forename: H L
authors.initials: HL
authors.lastname: Malech
mesh_terms: D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34; D001402:B-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D003573:Cytochrome b Group / Q000235:genetics*; D015316:Genetic Therapy; D006105:Granulomatous Disease, Chronic / Q000235:genetics* / Q000628:therapy; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics*; D019255:NADPH Oxidases; D013481:Superoxides / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D003573:Cytochrome b Group; D013481:Superoxides; C036888:cytochrome b558; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases