Ultrastructural characterization of cystic fibrosis sputum using atomic force and scanning electron microscopy
Ultrastructural characterization of cystic fibrosis sputum using atomic force and scanning electron microscopy
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DOI：
10.1016/j.jcf.2011.09.008
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发表时间：
2012-03-01
影响因子：
5.2
通讯作者：
Hart, Dominik
中科院分区：
文献类型：
--
作者：
Manzenreiter, Reinhard;Kienberger, Ferry;Hart, Dominik
背景：囊性纤维化(CF)肺疾病的特征是持续性中性粒细胞炎症和进行性组织破坏。中性粒细胞是CF气道液中的主要细胞成分，已知在刺激时会形成中性粒细胞胞外陷阱(Net)。CF气道液中存在大量细胞外DNA-Net。然而，Net对CF气道液基质成分的结构贡献仍然知之甚少。我们假设CF气道液由与亚细胞结构相关的不同DNA-Net组成。方法/主要发现：我们使用原子力显微镜(AFM)和扫描电子显微镜对CF痰的性质和Net在细胞外CF痰基质中的作用进行了超微结构表征。这些研究表明，CF痰主要由高密度的Net网络和NETsis衍生物质组成。不同脱氧核糖核酸酶作用于CF痰，可有效降解CFNet，有效液化CF痰中的粘液样结构。AFM结果定量分析显示，CF痰中存在3个球形组分，其中较大的两个组分具有中性粒细胞胞外体的特征。结论/意义：这些研究表明，过度的网状结构是导致CF痰凝胶状结构的主要因素，并为CF-Net含有外胚体样结构提供证据，可作为未来治疗的靶点。(C)2011年欧洲囊性纤维化协会。爱思唯尔出版，版权所有。
Background: Cystic fibrosis (CF) lung disease is characterized by perpetuated neutrophilic inflammation with progressive tissue destruction. Neutrophils represent the major cellular fraction in CF airway fluids and are known to form neutrophil extracellular traps (NETs) upon stimulation. Large amounts of extracellular DNA-NETs are present in CF airway fluids. However, the structural contribution of NETs to the matrix composition of CF airway fluid remains poorly understood. We hypothesized that CF airway fluids consist of distinct DNA-NETs that are associated to subcellular structures.Methodology/principal findings: We employed atomic force microcopy (AFM) and scanning electron microcopy to ultrastructurally characterize the nature of CF sputum and the role of NETs within the extracellular CF sputum matrix. These studies demonstrate that CF sputum is predominantly composed of a high-density meshwork of NETs and NETosis-derived material. Treatment of CF sputum with different DNases degraded CF NETs and efficiently liquefied the mucous-like structure of CF sputum. Quantitative analysis of AFM results showed the presence of three globular fractions within CF sputum and the larger two ones featured characteristics of neutrophil ectosomes.Conclusions/significance: These studies suggest that excessive NET formation represents the major factor underlying the gel-like structure of CF sputum and provide evidence that CF-NETs contain ectosome-like structures that could represent targets for future therapeutic approaches. (C) 2011 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.