Neutrophil extracellular traps (NET) induced by different stimuli: A comparative proteomic analysis
Neutrophil extracellular traps (NET) induced by different stimuli: A comparative proteomic analysis
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DOI：
10.1371/journal.pone.0218946
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发表时间：
2019-07-08
期刊：
影响因子：
3.7
通讯作者：
Migliorini, Paola
中科院分区：
文献类型：
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作者：
Petretto, Andrea;Bruschi, Maurizio;Migliorini, Paola
中性粒细胞胞外陷阱（NET）的形成是中性粒细胞对感染反应的一部分，但过度或不适当的NETosis可能引发自身抗体的产生并导致自身免疫性疾病的器官损伤。中性粒细胞自发网状并不常见，为了研究它们的结构，有必要在体外通过选定的刺激诱导NET。本研究采用蛋白质组学分析的方法研究了不同刺激下产生的NET的蛋白质组成和翻译后修饰。用PMA、A23187、大肠杆菌LPS或未经处理刺激健康供者的中性粒细胞；3小时后，清洗细胞，用dna酶和收集的上清进行质谱分析。数据分析采用无监督分层聚类分析。我们发现了任何来源或不受任何刺激的NET中包含的蛋白质：lps诱导的和自发的NET在蛋白质组成上存在差异，而PMA-和a23187诱导的NET似乎更相似。在我们研究的翻译后修饰中，蛋氨酸亚砜化是常见的，特别是在PMA和lps诱导的NETs中。髓过氧化物酶是更广泛修饰的蛋白质。因此，蛋白质组学分析表明，不同刺激诱导的NET在蛋白质组成和翻译后修饰方面存在异质性，这表明在不同条件下诱导的NET可能具有不同的生物学效应。
Neutrophil extracellular traps (NET) formation is part of the neutrophil response to infections, but excessive or inappropriate NETosis may trigger the production of autoantibodies and cause organ damage in autoimmune disorders. Spontaneously netting neutrophils are not frequent and induction of NET in vitro by selected stimuli is necessary to investigate their structure. In the present work, the protein composition and post-translational modifications of NET produced under different stimuli have been studied by means of proteomic analysis. Neutrophils from healthy donors were stimulated by PMA, A23187, Escherichia coli LPS or untreated; after three hours, cells were washed, treated with DNase and supernatants collected for mass spectrometry. Data were analyzed by unsupervised hierarchical clustering analyses. We identified proteins contained in NETs of any source or exclusive of one stimulus: LPS-induced and spontaneous NET diverge in protein composition, while PMA- and A23187-induced NET appear more similar. Among the post-translational modifications we examined, methionine sulfoxidation is frequent especially in PMA- and LPS-induced NETs. Myeloperoxidase is the protein more extensively modified. Thus, proteomic analysis indicates that NETs induced by different stimuli are heterogeneous in terms of both protein composition and post-translational modifications, suggesting that NET induced in different conditions may have different biological effects.