Genetic Correction of p67 Deficient Chronic Granulomatous Disease Using Peripheral Blood Progenitor Cells as a Target for Retrovirus Mediated Gene Transfer

Chronic granulomatous disease (CGD) can result from any from transduced CD34 progenitors from the p67 CGD patient were oxidase positive with the average level of corof four single gene defects involving the components of the superoxide (O2 ) generating phagocyte nicotinamide aderection per granulocyte of 85% of that seen with granulocytes in similar cultures of CD34 progenitors from normal nine dinucleotide phosphate (NADPH) oxidase. We show that transduction of peripheral blood CD34 hematopoietic volunteers. Nitroblue tetrazolium dye reduction assays of colonies of transduced progenitors in soft agar indicated progenitors from a p67 deficient CGD patient with replication defective amphotropic retrovirus encoding p67 that in some studies restoration of oxidase activity occurred in myeloid cells within 44% of granulocyte-erythrocyte(MFGS-p67) significantly corrected the CGD functional defect in phagocyte oxidase activity in vitro. Using a chemimonocyte colonies, and within 28% of the combined group of granulocyte colonies/monocyte colonies/granulocyteluminescence assay of oxidase activity, we showed that transduced patient CD34 progenitors differentiating to mymonocyte colonies. These high correction rates were achieved without any selective regimen to enrich for transeloid cells in culture produced 25% of the total superoxide produced by normal CD34 progenitors differentiating in culduced cells. This study provides a basis for development of gene therapy for the p67 deficient form of CGD. ture. A flow cytometric assay of oxidase activity used to assess the oxidase function of individual cells in the cultures This is a US government work. There are no restrictions on its use. indicated that up to 32% of maturing granulocytes derived