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 × 105 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 (≤ 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. © 1994 by The American Society of Hematology.