Hassan N F, Campbell D E, Douglas S D
Clin Immunol Immunopathol. 1987 Mar;42(3):274-80. doi: 10.1016/0090-1229(87)90015-8.
O-Phenylenediamine (OPD) oxidation has been extensively utilized for the measurement of peroxidase-mediated catabolism of hydrogen peroxide. However, until now this system has not been evaluated for the measurement of hydrogen peroxide produced upon activation of the hexose monophosphate shunt (HMPS) in polymorphonuclear leukocytes (PMNs). OPD oxidation by phorbol myristate acetate (PMA)-stimulated PMNs was mediated by both hydrogen peroxide and superoxide produced by the activation of the HMPS. Furthermore, OPD oxidation by an oxidative mechanism independent of the HMPS was observed by the PMA stimulation of PMNs obtained from patients with chronic granulomatous disease (CGD). This HMPS-independent OPD oxidation was inhibited by superoxide dismutase or 1 mM potassium cyanide (KCN). Superoxide dismutase, catalase, or 1 mM potassium cyanide inhibited 50% OPD oxidation obtained with PMA-stimulated normal PMNs. PMA treatment of purified human myeloperoxidase (MPO) produced OPD oxidation which is inhibited by superoxide dismutase or 1 mM KCN. These data indicate that OPD oxidation observed with CGD PMNs is mediated by a PMA-induced oxidase activity of myeloperoxidase. OPD oxidation in the presence of 1 mM KCN is a method comparable in sensitivity with ferricytochrome c reduction for the evaluation of HMPS activity. Furthermore, the OPD assay can measure myeloperoxidase oxidase activity in PMA-stimulated PMNs.
邻苯二胺（OPD）氧化已被广泛用于测量过氧化物酶介导的过氧化氢分解代谢。然而，到目前为止，该系统尚未用于测量多形核白细胞（PMN）中己糖磷酸旁路（HMPS）激活时产生的过氧化氢。佛波酯（PMA）刺激的PMN对OPD的氧化是由HMPS激活产生的过氧化氢和超氧化物介导的。此外，通过PMA刺激慢性肉芽肿病（CGD）患者的PMN，观察到一种独立于HMPS的氧化机制导致的OPD氧化。这种独立于HMPS的OPD氧化被超氧化物歧化酶或1 mM氰化钾（KCN）抑制。超氧化物歧化酶、过氧化氢酶或1 mM氰化钾抑制了PMA刺激的正常PMN产生的50%的OPD氧化。用PMA处理纯化的人髓过氧化物酶（MPO）会产生OPD氧化，该氧化被超氧化物歧化酶或1 mM KCN抑制。这些数据表明，CGD患者的PMN中观察到的OPD氧化是由PMA诱导的髓过氧化物酶氧化酶活性介导的。在1 mM KCN存在下的OPD氧化是一种在灵敏度上与铁细胞色素c还原相当的用于评估HMPS活性的方法。此外，OPD测定法可以测量PMA刺激的PMN中的髓过氧化物酶氧化酶活性。
