Borregaard N
Immunol Lett. 1985;11(3-4):165-71. doi: 10.1016/0165-2478(85)90164-6.
Using nitrogen cavitation and Percoll density gradient centrifugation for subcellular centrifugation of human neutrophils, approximately 90% of the low potential b-cytochrome, unique for phagocytes, as well as 50% of the flavoproteins in normal neutrophils were found in a granule fraction which co-sedimented with the specific granules. Upon stimulation of the intact cells with phorbol myristate acetate, both the b-cytochrome and the flavoprotein translocated from this granule fraction to the fractions which contained the plasma membranes and the NADPH oxidase activity. In neutrophils from two patients with chronic granulomatous disease, both the b-cytochrome and the flavoprotein of the granules were absent, but flavoprotein was present in normal amounts in the membrane and cytosol fractions. Taken together, these findings suggest that the specific granules, or granules co-sedimenting with the specific granules, are important stores for the components of the NADPH oxidase, which is responsible for the respiratory burst. Analysis of the stoichiometry of CO2 generation, H+ secretion and O2 consumption by stimulated neutrophils indicated that the hexose monophosphate shunt is the source of both protons and electrons for the NADPH oxidase activity, as well as of the extra protons secreted during the respiratory burst.
利用氮气空化和Percoll密度梯度离心法对人中性粒细胞进行亚细胞离心，发现吞噬细胞特有的约90%的低电位b - 细胞色素以及正常中性粒细胞中50%的黄素蛋白存在于与特异性颗粒共同沉降的颗粒组分中。用佛波酯刺激完整细胞后，b - 细胞色素和黄素蛋白都从该颗粒组分转移到含有质膜和NADPH氧化酶活性的组分中。在两名慢性肉芽肿病患者的中性粒细胞中，颗粒中的b - 细胞色素和黄素蛋白均缺失，但膜和胞质组分中的黄素蛋白含量正常。综上所述，这些发现表明特异性颗粒或与特异性颗粒共同沉降的颗粒是NADPH氧化酶组分的重要储存部位，NADPH氧化酶负责呼吸爆发。对受刺激的中性粒细胞产生二氧化碳、分泌氢离子和消耗氧气的化学计量分析表明，磷酸己糖旁路是NADPH氧化酶活性所需质子和电子的来源，也是呼吸爆发期间分泌的额外质子的来源。
