A Determination of H2O2 Release by the Treatment of HumanBlood Polymorphonuclear Leukocytes with Myristate

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Abstract
Free H2O2 released from human blood leukocytes during phagocytosis into the extracellular medium was highly reactive with the ferric form of HRP, forming an enzyme-substrate complex which was identical to HRP-H2O2 compound II. The formation of HRP-H2O2 compound II was employed for assaying the rates of H2O2 release by leukocytes upon addition of bacteria or myristate. The treatment of normal human blood leukocytes with myristate resulted in a marked stimulation of H2O2 release compared to phagocytizing cells. The activity of H2O2 release in response to myristate was found to be deficient in the leukocytes of two patients with chronic granulomatous disease. This assay method with myristate supplementation is so sensitive and specific that it should be useful for the diagnosis of chronic granulomatous disease.