Mouse Neutrophil Extracellular Traps in Microbial Infections
Mouse Neutrophil Extracellular Traps in Microbial Infections
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DOI：
10.1159/000205281
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发表时间：
2009-01-01
影响因子：
5.3
通讯作者：
Brinkmann, Volker
中科院分区：
文献类型：
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作者：
Ermert, David;Urban, Constantin F.;Brinkmann, Volker
神经细胞外陷阱（NETs）在机体对微生物感染的天然免疫中起重要作用。NET已经在几个物种中描述，但NET形成的分子细节及其在感染中的作用尚未得到解决，部分原因是我们缺乏最佳的实验模型。在这里，我们描述的工具，调查NET形成从小鼠中分离的中性粒细胞。在用PMA体外刺激野生型小鼠中性粒细胞后，我们分析了NET形成过程中的3个重要步骤：活性氧（ROS）产生、NET细胞死亡和NET释放。正如预期的那样，来自NADPH氧化酶缺陷小鼠的中性粒细胞不能产生ROS，并且在刺激后既不死亡也不释放NET。我们发现，从几个小鼠品系的中性粒细胞产生NET的效率不同，NET的形成与ROS的产生量相关。用白色念珠菌激活也导致ROS产生和NET细胞死亡。该真菌的菌丝形式比酵母形式更有效地诱导NETs。通过这项工作，我们提供了工具，研究在体外NET组装在鼠标系统。版权所有（C）2009 S，Karger AG，巴塞尔
Neutrophil extracellular traps (NETs) play an important role in innate immunity to microbial infections. NETs have been described in several species, but the molecular details of NET formation and their role in infection has not been addressed, partly because we lack optimal experimental models. Here we describe tools to investigate NET formation in neutrophils isolated from mice. Upon in vitro stimulation of wildtype mouse neutrophils with PMA, we analyzed 3 important steps in the process of NET formation: reactive oxygen species (ROS) production, NET cell death and NET release. As expected, neutrophils from NADPH oxidase-deficient mice failed to produce ROS and did not die nor release NETs upon stimulation. We found that neutrophils from several mouse strains produced NETs with different efficiency and that NET formation correlated with the amount of ROS produced. Activation with Candida albicans also resulted in ROS production and NET cell death. The hyphal form of this fungus induced NETs more effectively than the yeast form. With this work, we provide tools to study in vitro NET assembly in the mouse system. Copyright (C) 2009 S, Karger AG, Basel