A Label-Free Quantitative Proteomic Analysis of Mouse Neutrophil Extracellular Trap Formation Induced by Streptococcus suis or Phorbol Myristate Acetate (PMA)
A Label-Free Quantitative Proteomic Analysis of Mouse Neutrophil Extracellular Trap Formation Induced by Streptococcus suis or Phorbol Myristate Acetate (PMA)
复制标题
对猪链球菌或肉豆蔻酸佛波醇醋酸酯 (PMA) 诱导的小鼠中性粒细胞胞外陷阱形成的无标记定量蛋白质组学分析
DOI：
10.3389/fimmu.2018.02615
复制
发表时间：
2018-11-13
影响因子：
7.3
通讯作者：
Han，Li
中科院分区：
文献类型：
--
作者：
Wang，Xiaoping;Zhao，Jianqing;Han，Li
猪链球菌(S.suis)是世界范围内最重要的五种猪病原体之一，有时会威胁人类健康，特别是在与猪或猪肉制品有密切接触的人中。猪链球菌感染可在体外和体内诱导中性粒细胞胞外陷阱(Net)的形成，而中性粒细胞胞外陷阱(Net)的结构对猪链球菌的清除起着至关重要的作用。然而，猪链球菌诱导Net形成的信号通路仍有待阐明。在本研究中，我们使用了一种无标记的定量蛋白质组学分析方法，对猪链球菌或佛波酯(PMA)诱导的小鼠网络形成进行了研究。在感染猪链球菌的中性粒细胞中，超过50%的差异表达蛋白显示出与PMA刺激后相似的变化，这两种刺激诱导的网状形成需要PKC、NADPH氧化酶和MPO。由于PMA能诱导较强的网络形成，而猪链球菌(MOI=2)只诱导较弱的网络形成，因此诱导剂与网络形成之间的关系值得考虑。有趣的是，参与多肽酶活性的蛋白质对每种诱导剂的反应表现出显著的差异。在这些多肽酶中，MMP8的表达对PMA有明显的抑制作用，而对猪链球菌无明显影响。随后的一项研究进一步证实，基质金属蛋白酶-8的活性与这两种刺激诱导的网络形成呈负相关。因此，本研究提供了有关中性粒细胞对诱导剂反应形成Net的方式的潜在重要信息。
Streptococcus suis (S. suis) ranks among the five most important porcine pathogens worldwide and occasionally threatens human health, particularly in people who come into close contact with pigs or pork products. An S. suis infection induces the formation of neutrophil extracellular traps (NETs) in vitro and in vivo, and the NET structure plays an essential role in S. suis clearance. However, the signaling pathway by which S. suis induces NET formation remains to be elucidated. In the present study, we used a label-free quantitative proteomic analysis of mouse NET formation induced by S. suis or phorbol myristate acetate (PMA), a robust NET inducer. Greater than 50% of the differentially expressed proteins in neutrophils infected by S. suis showed similar changes as observed following PMA stimulation, and PKC, NADPH oxidase, and MPO were required for NET formation induced by both stimuli. Because PMA induced robust NET formation while S. suis (MOI = 2) induced only weak NET formation, the association between the inducer and NET formation was worth considering. Interestingly, proteins involved in peptidase activity showed significant differential changes in response to each inducer. Of these peptidases, MMP-8 expression was obviously decreased in response to PMA, but it was not significantly changed in response to S. suis. A subsequent study further confirmed that MMP-8 activity was inversely correlated with NET formation induced by both stimuli. Therefore, the present study provides potentially important information about the manner by which neutrophils responded to the inducers to form NETs.