Abstract
In our study, we investigated the intracellular killing ability of granulocytes for healthy and ill subjects by measuring NADH oxidase activity and release of hydrogen peroxide. The protocol methodology measured the hydrogen peroxide released after granulocytes activation by PMA (phorbol 12-myristate 13-acetate) by using the Amplex Red assay, which included counting granulocytes by flow cytometer and measurement of the kinetic curve of NADPH oxidase activity by fluorometer. Two parameters were used to describe the level of granulocyte activity: the initial rate of NADPH-oxidase enzyme activity and the level of hydrogen peroxide released after 20 min of granulocyte activation. The method was applied to measure granulocyte activity in 55 healthy subjects and 30 patients with cancer. It was demonstrated that applied procedure is sensitive for estimation of the disease activity. The granulocyte activity in patients with cancer was compared with the granulocyte activity of healthy subjects and demonstrated the down-regulation of NADPH oxidase activity. We showed that granulocytes of cancer patients had inhibited oxidative burst and less NADPH oxidase activity.
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