Proteomic Characterization of Middle Ear Fluid Confirms Neutrophil Extracellular Traps as a Predominant Innate Immune Response in Chronic Otitis Media.
Proteomic Characterization of Middle Ear Fluid Confirms Neutrophil Extracellular Traps as a Predominant Innate Immune Response in Chronic Otitis Media.
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DOI：
10.1371/journal.pone.0152865
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Preciado D
中科院分区：
文献类型：
--
作者：
Val S;Poley M;Brown K;Choi R;Jeong S;Colberg-Poley A;Rose MC;Panchapakesan KC;Devaney JC;Perez-Losada M;Preciado D
慢性中耳炎（COM）的特征是中耳积液（中耳积液）和传导性听力损失。这反映了粘液分泌过多，但粘膜组分的整体蛋白质组学分析是有限的。本研究旨在表征COM儿童MEE的蛋白质组，目的是阐明重要的先天免疫反应。从接受鼓膜切开术的COM儿童（n = 49）中收集MEE。采用质谱法对9个样品进行蛋白质组学分析。通过细胞因子多重测定、免疫印迹、中性粒细胞弹性蛋白酶活性、下一代DNA测序和/或免疫荧光分析进一步分析独立样品。通过MS鉴定了109种独特和常见的蛋白质。大多数是先天免疫分子，沿着典型的细胞内蛋白质，如组蛋白和肌动蛋白。所有映射的肽计数的19.5%来自已知由中性粒细胞释放的蛋白质。免疫荧光和免疫印迹证明每个MEE中存在中性粒细胞细胞外陷阱（NET），并沿着MUC 5 B共定位。在渗出液中发现的DNA显示了人类起源的完整DNA。MEE的蛋白质组学分析揭示了一种主要的嗜酸性先天性粘膜反应，其中MUC 5 B与NET DNA相关。NET是人COM中耳积液的主要大分子成分。
Chronic Otitis Media (COM) is characterized by middle ear effusion (MEE) and conductive hearing loss. MEE reflect mucus hypersecretion, but global proteomic profiling of the mucosal components are limited. This study aimed at characterizing the proteome of MEEs from children with COM with the goal of elucidating important innate immune responses. MEEs were collected from children (n = 49) with COM undergoing myringotomy. Mass spectrometry was employed for proteomic profiling in nine samples. Independent samples were further analyzed by cytokine multiplex assay, immunoblotting, neutrophil elastase activity, next generation DNA sequencing, and/or immunofluorescence analysis. 109 unique and common proteins were identified by MS. A majority were innate immune molecules, along with typically intracellular proteins such as histones and actin. 19.5% percent of all mapped peptide counts were from proteins known to be released by neutrophils. Immunofluorescence and immunoblotting demonstrated the presence of neutrophil extracellular traps (NETs) in every MEE, along with MUC5B colocalization. DNA found in effusions revealed unfragmented DNA of human origin. Proteomic analysis of MEEs revealed a predominantly neutrophilic innate mucosal response in which MUC5B is associated with NET DNA. NETs are a primary macromolecular constituent of human COM middle ear effusions.