A new genetic subgroup of chronic granulomatous disease with autosomal recessive mutations in p40^phox and selective defects in neutrophil NADPH oxidase activity

Chronic granulomatous disease (CGD), an immunodeficiency with recurrent pyogenic infections and granulomatous in-flammation, results from loss of phagocyte superoxide production by recessive mutations in any 1 of 4 genes encoding subunits of the phagocyte NADPH oxidase. These include gp91 ^phox and p22_phox, which form the membrane-integrated flavocytochrome b, and cytosolic subunits p47^phox and p67 ^phox. A fifth subunit, p40^phox, plays an important role in phagocytosis-induced superoxide production via a phox homology (PX) domain that binds to phosphatidylinositol 3-phosphate (PtdIns(3)P). We report the first case of autosomal recessive mutations in NCF4, the gene encoding p40 ^phox, in a boy who presented with granulomatous colitis. His neutrophils showed a substantial defect in intracellular superoxide production during phagocytosis, whereas extracellular release of superoxide elicited by phorbol ester or formyl-methionyl-leucyl-phenylalanine (fMLF) was unaffected. Genetic analysis of NCF4 showed compound heterozygosity for a frameshift mutation with premature stop codon and a missense mutation predicting a R105Q substitution in the PX domain. Parents and a sibling were healthy heterozygous carriers. p40^phoxR105Q lacked binding to PtdIns(3)P and failed to reconstitute phagocytosis-induced oxidase activity in p40^phox- deficient granulocytes, with premature loss of p40^phoxR105Q from phagosomes. Thus, p40^phox binding to PtdIns(3)P is essential for phagocytosis-induced oxidant production in human neutrophils and its absence can be associated with disease.