title: Retrovirus-mediated reconstitution of respiratory burst activity in X-linked chronic granulomatous disease cells.
abstract: X-linked chronic granulomatous disease (X-CGD) results from mutations in the gene encoding gp91phox, the larger subunit of the respiratory burst oxidase cytochrome b. In this study, a recombinant retrovirus vector was constructed and evaluated for its expression of human gp91phox in a human X-CGD myeloid cell line in which the endogenous gp91phox gene had been disrupted by gene targeting. The retrovirus construct, Zip/PGKgp91, was first introduced into the GP+envAm12 amphotropic packaging line and yielded virus producer clones with estimated titers of up to 1 x 10(5) cfu/mL. Coculture infection of X-CGD myeloid cells with Zip/PGKgp91 resulted in restoration of respiratory burst activity to 15% of the cells. Isolated clonal infectants expressed relatively low levels of recombinant gp91phox (< or = 12% of wild-type), but exhibited considerable superoxide-generating activity (up to nearly 60% of wild-type). These results show the feasibility of phenotypic correction of CGD using gene replacement therapy and suggest that even modest levels of gp91phox expression may lead to considerable functional correction of X-CGD neutrophils.
authors.affiliation: Department of Pediatrics, Herman B Wells Center for Pediatric Research, James Whitcomb Riley Hospital for Children, Indiana University Medical Center, Indianapolis 46202-5225.
authors.forename: A
authors.initials: A
authors.lastname: Kume
authors.forename: M C
authors.initials: MC
authors.lastname: Dinauer
mesh_terms: D015152:Blotting, Northern; D002454:Cell Differentiation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002999:Clone Cells; D015786:Cytochromes b5 / Q000096:biosynthesis / Q000235:genetics; D005822:Genetic Vectors; D006098:Granulocytes / Q000166:cytology / Q000378:metabolism; D006105:Granulomatous Disease, Chronic / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics; D009154:Mutation; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D016897:Respiratory Burst; D012190:Retroviridae; D013481:Superoxides / Q000378:metabolism; D014162:Transfection; D014960:X Chromosome
chemical_list: D008562:Membrane Glycoproteins; D013481:Superoxides; D015786:Cytochromes b5; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases