Impaired production of reactive oxygen species (ros) in glucose-6-phosphate dehydrogenase (g6pd) deficient granulocytes

Neutrophil dysfunction and increased susceptibility to infection have been reported in severe G6PD deficiency. The detail mechanism has not been fully elucidated. In the current study, we compared the production of ROS by G6PDdeficient granulocytes to that by normal controls after the addition of phorbol myristate acetate (PMA) as stimulant. Leukocytes were isolated by Histopaque method. Production of H₂O₃ was measured by a fluorescent method. In this method, non-fluorescent 4-hydroxphenylacetic acid was converted to fluorescent e.e'-dihydroxy-(1, 1'-bipheny)-3, 3' diacetic acid in the presence of horse radish peroxidase and H₂O₂- Superoxide production was monitored using this same method with the addition of Superoxide dismutase which immediately converted Superoxide to H₂O₂. The production of Superoxide and H₂O₂ was singificantly reduced in G6PD-deficient leukocytes (2.4±0.36 nmole/10⁵ granulocytes & 2.26 ± 0.95 per10⁵, respectively) vs controls(0.90±0.36 nmole/10⁵granulocytes & 0.80±0.36 nmole/10⁵granulocytes, repectively). We also determined Superoxide production by granulocytes in the presence PMA using a ultra-sensitive chemiluminescence method with lucigenin amplification. The result obtained by this method is consistent with the fluorescent method. Taken toghether, these data indicate that decreased production of ROS contributes to neutrophil dysfunction in G6PD-deficiency.