Production of hydrogen peroxide by phagocytizing human granulocytes.
Production of hydrogen peroxide by phagocytizing human granulocytes.
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通过吞噬人粒细胞产生过氧化氢。
DOI：
10.1016/0304-4165(68)90116-5
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
Dirk Roos
中科院分区：
文献类型：
--
作者：
Homan;R. S. Weening;Dirk Roos
本文报道了一种快速、灵敏的定量测定人粒细胞吞噬产生的H_2O_2的方法。为此，Keston和Brandt的方法被机械化，其基于在过氧化物酶存在下通过H2 O2将非荧光的白细胞乙酰基-2，7-二氯荧光素氧化成荧光化合物。确定了该试验的最佳条件。水中的H2 O2可以在0.05至0.5 μ M的范围内测量，在0.4 μ M时标准偏差为1.2%（n = 1-）。通过吞噬粒细胞产生的H2 O2只能在含有磷酸盐缓冲盐、白蛋白、葡萄糖、NaN 3和IgG包被的乳胶颗粒的培养基中测量。荧光信号是过氧化氢酶敏感的。在已知量的H2 O2中，添加到该介质中，回收了97%。在最佳条件下，我们发现每小时每10-10个细胞（10个不同的健康供体）产生970 ± 170 mumoles的H2 O2，相当于观察到的O2消耗增加的50%至70%。2例慢性肉芽肿病患者的吞噬粒细胞不产生H2 O2，而杂合子的吞噬粒细胞产生中等水平的H2 O2。
A rapid and sensitive method is described for the quantitative determination of H202 produced by phagocytizing human granulocytes. For this purpose, the method of Keston and Brandt was mechanized, which is based on the oxidation of nonfluorescent leukociacetyl-2,7-dichlorofluorescein to a fluorescent compound by H202 in the presence of peroxidase. The optimal conditions for this test were determined. H202 in water can be measured in the range of 0.05 to 0.5 muM, with a standard deviation of 1.2 per cent at 0.4 muM (n = 1-). The production of H2O2 by phagocytizing granulocytes could only be measured in a medium which contained phosphate-buffered salt, albumin, glucose, NaN3, and IgG-coated latex particles. The fluorescence signal was catalase-sensitive. Of known amounts of H202, added to this medium, 97 per cent were recovered. Under optimal conditions we found a H2O2 production of 970 plus or minus 170 mumoles per 10-10 cells per hour (10 different healthy donors), corresponding to 50 to 70 per cent of the observed increase in O2 consumption. No H2O2 was produced by phagocytizing granulocytes from 2 patients with chronic granulomatous disease, while intermediate values were found in the cells from heterozygotes.