REEVALUATION OF CYTOCHROME B AND FLAVIN ADENINE-DINUCLEOTIDE IN NEUTROPHILS FROM PATIENTS WITH CHRONIC GRANULOMATOUS-DISEASE AND DESCRIPTION OF A FAMILY WITH PROBABLE AUTOSOMAL RECESSIVE INHERITANCE OF CYTOCHROME-B DEFICIENCY

  • 1 April 1986
    • journal article
    • research article
    • Vol. 67  (4) , 1132-1138
Abstract
Chronic granulomatous disease (CGD) is a genetically heterogeneous syndrome characterized by a microbial killing defect of polymorphonuclear leukocytes (PMNs) due to lack of superoxide .**GRAPHIC**. generation. Recent studies indicate that the neutrophil .**GRAPHIC**. system consists of at least two components, flavoprotien-flavin adenine dinucleotide (FAD)- and cytochrome b. We evaluate the cytochrome b and FAD content in PMN from 30 CGD patients. The method for quantitating cytochrome b was modified by using PMN sonicates incubated with azide plus hydrogen peroxide. With this approach, several absorption peaks corresponding to myeloperoxidase and eosinophil peroxidase, which overlap with peaks of cytochrome b, were obliterated from reduced-minus-oxidized spectra, whereas the peaks of cytochrome b were not and could be readily quantitated. Cytochrome b was detected in PMNs from all 24 normal adults (47.4 .+-. 2.9 pmol/7.5 .times. 106 cells), was absent in PMNs from 11 male CGD patients and one female CGD patient, but was present in normal amounts in PMNs from nine male and nine female CGD patients. Stimulated nitroblue tetrazolium (NBT) tests performed on PMNs from mothers of CGD patients indicated that cytochrome b deficiency was associated with X-linked inheritance, except in one case in which probable autosomal recessive inheritance was demonstrated. The PMN NBT test of the mother of another male patient without cytochrome b deficiency suggested an X-linked form of inheritance. In related studies, the FAD content in PMN particulate fractions was reduced in 4 of 28 CGD patients studied. All four CGD patients with reduced FAD lacked cytochrome b. However, three patients with cytochrome b deficiency had normal FAD. Thus, the results indicate that PMN cytochrome b deficiency is observed in most X-linked and in some autosomal recessive CGD, that cytochrome b deficiency may be associated with FAD deficiency, and that cytochrome b and FAD are normal in most patients with non-X-linked CGD.