Kenny Elaine F, Herzig Alf, Krüger Renate, Muth Aaron, Mondal Santanu, Thompson Paul R, Brinkmann Volker, Bernuth Horst von, Zychlinsky Arturo
Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany.
Department of Paediatric Pneumology and Immunology, Outpatient Clinic for Primary Immunodeficiencies, Charité Medical School, Berlin, Germany.
Elife. 2017 Jun 2;6:e24437. doi: 10.7554/eLife.24437.
Neutrophils release neutrophil extracellular traps (NETs) which ensnare pathogens and have pathogenic functions in diverse diseases. We examined the NETosis pathways induced by five stimuli; PMA, the calcium ionophore A23187, nigericin, and Group B Streptococcus. We studied NET production in neutrophils from healthy donors with inhibitors of molecules crucial to PMA-induced NETs including protein kinase C, calcium, reactive oxygen species, the enzymes myeloperoxidase (MPO) and neutrophil elastase. Additionally, neutrophils from chronic granulomatous disease patients, carrying mutations in the NADPH oxidase complex or a MPO-deficient patient were examined. We show that PMA, and GBS use a related pathway for NET induction, whereas ionophores require an alternative pathway but that NETs produced by all stimuli are proteolytically active, kill bacteria and composed mainly of chromosomal DNA. Thus, we demonstrate that NETosis occurs through several signalling mechanisms, suggesting that extrusion of NETs is important in host defence.
中性粒细胞释放中性粒细胞胞外诱捕网（NETs），其可捕获病原体并在多种疾病中发挥致病作用。我们研究了由五种刺激物诱导的NETosis途径，即佛波酯（PMA）、钙离子载体A23187、尼日利亚菌素和B族链球菌。我们使用对PMA诱导的NETs至关重要的分子抑制剂，研究了健康供体中性粒细胞中的NET生成，这些分子包括蛋白激酶C、钙、活性氧、髓过氧化物酶（MPO）和中性粒细胞弹性蛋白酶。此外，还检测了慢性肉芽肿病患者的中性粒细胞，这些患者的NADPH氧化酶复合物存在突变，以及一名MPO缺陷患者的中性粒细胞。我们发现，PMA和B族链球菌使用相关途径诱导NETs，而离子载体则需要另一种途径，但所有刺激物产生的NETs都具有蛋白水解活性，能杀死细菌，且主要由染色体DNA组成。因此，我们证明NETosis通过多种信号传导机制发生，这表明NETs的排出在宿主防御中很重要。
