FUNCTIONAL RECONSTITUTION OF THE NADPH-OXIDASE BY ADENOASSOCIATED VIRUS GENE-TRANSFER

Chronic granulomatous disease (CGD) comprises a heterogeneous group of inherited conditions characterized biochemically by disordered functi on of a unique multicomponent enzyme system present in phagocytic cell s, the NADPH-oxidase. Clinically, it is characterized by recurrent bac terial and fungal infections that are relatively resistant to treatmen t by conventional means, Curative bone marrow transplantation has been successfully achieved in a small number of cases, but the wider appli cation of this procedure is limited by availability of suitable donor material. Somatic gene therapy would overcome this problem, and severa l groups have now shown correction of the biochemical defect in hemato poietic cells by retrovirus-mediated gene transfer. However, the failu re of the current generation of retroviral vectors to efficiently tran sduce quiescent cells greatly restricts their potential for gene trans fer to pluripotent hematopoietic stem cells. Given these limitations, we have constructed vectors based on adeno-associated virus and used t hese to transfer a functional copy of the p47(phox) gene to immortaliz ed B cells derived from patients with p47(phox)-deficient autosomal re cessive CGD. We show stable expression of protein and restoration of N ADPH-oxidase function in these cells in the absence of selection. Aden oassociated virus vectors may overcome some of the limitations of retr oviral gene delivery systems and may therefore be a useful vehicle for curative gene therapy of CGD and other primary immunodeficiencies. (C ) 1995 by The American Society of Hematology.