The induction of eosinophil peroxidase release: improved methods of measurement and stimulation
The induction of eosinophil peroxidase release: improved methods of measurement and stimulation
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DOI：
10.1016/j.jim.2004.05.003
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发表时间：
2004-08-01
影响因子：
2.2
通讯作者：
Moqbel, R
中科院分区：
文献类型：
--
作者：
Adamko, DJ;Wu, YQ;Moqbel, R
生产。和嗜酸性粒细胞过氧化物酶（EPO）的释放。与人类病理学有关。使用嗜酸性粒细胞进行脱颗粒测定通常非常困难，释放值非常低。 EPO 因其高阳离子电荷而独特。因此，它粘附在大多数细胞外表面，使得与其他释放的细胞蛋白相比更难以测量。基于对 EPO 粘性性质的了解，我们担心无法在受刺激细胞的上清液中重复测量体外释放的 EPO。相反，我们怀疑大部分释放的 EPO 粘附在管壁上。我们选择使用过氧化物酶底物邻苯二胺 (OPD) 来研究 EPO 活性的测量。与其他过氧化物酶底物不同，OPD 可溶于生理水溶液，不会裂解细胞膜，因此我们可以将 OPD 直接添加到嗜酸性粒细胞中并专门测量细胞外 EPO。这种新颖的方法将消除由于其粘合性质而导致 EPO 测量不正确的担忧。此外，我们开发了这种方法来根据 EPO 浓度来量化 EPO 释放。最后，利用这项技术，我们已经能够证明分泌型 IgA (s-IgA) 诱导的 EPO 释放。通过使用 ORD，我们开发了一种更灵敏、更特异的方法来分析细胞外 EPO 的释放。 (C) 2004 Elsevier B.V. 保留所有权利。
The production. and release of eosinophil peroxidase (EPO) has. been associated with human pathology. Degranulation assays with eosinophils are typically very difficult to do, with very low release values. EPO is unique for its high cationic charge. As such, it adheres to most extracellular surfaces, rendering it more difficult to measure compared with other released cellular proteins. Based on the understanding of the sticky nature of EPO, we were concerned that EPO released in vitro cannot be reproducibly measured in the supernatants of stimulated cells. Instead, we suspected that much of the released EPO was left adherent to the tube walls. We chose to investigate the measurement of EPO activity using the peroxidase substrate, O-phenylenediamine (OPD). Unlike other peroxidase substrates, OPD is soluble in aqueous physiological solutions, which do not lyse cell membranes, thereby allowing us to add OPD directly to eosinophils and exclusively measure extracellular EPO. This novel approach would remove the concerns of incorrect EPO measurements due to its adhesive nature. In addition, we developed this method to quantify EPO release in terms of EPO concentration. Finally, using this technique, we have been able to demonstrate secretory IgA (s-IgA)-induced release of EPO. By using ORD, we have developed a more sensitive and specific method to analyze the release of extracellular EPO. (C) 2004 Elsevier B.V. All rights reserved.