Exosomes in the nose induce immune cell trafficking and harbour an altered protein cargo in chronic airway inflammation.
Exosomes in the nose induce immune cell trafficking and harbour an altered protein cargo in chronic airway inflammation.
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DOI：
10.1186/s12967-016-0927-4
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发表时间：
2016-06-20
影响因子：
7.4
通讯作者：
Lötvall J
中科院分区：
文献类型：
--
作者：
Lässer C;O'Neil SE;Shelke GV;Sihlbom C;Hansson SF;Gho YS;Lundbäck B;Lötvall J
外泌体是纳米尺寸的细胞外囊泡，参与健康和疾病中的细胞间通讯。然而，关于外泌体在上呼吸道中的功能和分子组成的知识是有限的。因此，本研究的目的是确定鼻外泌体是否可以影响炎症细胞，并在健康受试者中建立鼻灌洗液来源的外泌体的蛋白质组，以及其在慢性气道炎性疾病[哮喘和慢性鼻窦炎（CRS）]个体中的改变。收集14名健康受试者、15名哮喘受试者和13名哮喘/CRS受试者的鼻灌洗液。用差速离心分离外泌体，并通过LC-MS/MS应用两个排除列表以及使用定量蛋白质组学分析蛋白质组。使用免疫途径分析和GO术语查找器来预测与外泌体蛋白质组相关的功能，并且使用迁移测定来分析鼻外泌体对免疫细胞的影响。首先，我们证明了鼻外泌体可以在体外诱导几种免疫细胞，如单核细胞、中性粒细胞和NK细胞的迁移。其次，使用质谱法，应用排除列表，生成来自健康受试者的外泌体的全面蛋白质清单。排除列表的使用导致鉴定了约15%的额外蛋白质，并增加了约20%的已鉴定蛋白质的置信度。总共在鼻外泌体中鉴定了604种蛋白质，鼻外泌体蛋白质组显示出与免疫相关功能（如免疫细胞运输）的强烈关联。第三，使用定量蛋白质组学方法来确定作为气道炎性疾病的结果的外泌体蛋白质组的改变。与健康对照相比，呼吸系统疾病受试者的外泌体中血清相关蛋白和粘蛋白更丰富，而具有抗菌功能的蛋白和屏障相关蛋白的表达减少。鼻外泌体显示诱导先天免疫细胞的迁移，这可能是重要的，因为气道上皮是抵抗病原体和过敏原的第一道防线。在患有气道疾病的受试者中，屏障和抗微生物外泌体蛋白的表达降低可能有助于增加对感染的易感性，这在疾病进展中具有重要的临床意义。本文的在线版本（doi：10.1186/s12967-016-0927-4）包含补充材料，可供授权用户使用。
Exosomes are nano-sized extracellular vesicles participating in cell-to-cell communication both in health and disease. However, the knowledge about the functions and molecular composition of exosomes in the upper airways is limited. The aim of the current study was therefore to determine whether nasal exosomes can influence inflammatory cells and to establish the proteome of nasal lavage fluid-derived exosomes in healthy subjects, as well as its alterations in individuals with chronic airway inflammatory diseases [asthma and chronic rhinosinusitis (CRS)]. Nasal lavage fluid was collected from 14 healthy subjects, 15 subjects with asthma and 13 subjects with asthma/CRS. Exosomes were isolated with differential centrifugation and the proteome was analysed by LC–MS/MS with the application of two exclusion lists as well as using quantitative proteomics. Ingenuity Pathways Analysis and GO Term finder was used to predict the functions associated with the exosomal proteome and a migration assay was used to analyse the effect on immune cells by nasal exosomes. Firstly, we demonstrate that nasal exosomes can induce migration of several immune cells, such as monocytes, neutrophils and NK cells in vitro. Secondly, a mass spectrometry approach, with the application of exclusion lists, was utilised to generate a comprehensive protein inventory of the exosomes from healthy subjects. The use of exclusion lists resulted in the identification of ~15 % additional proteins, and increased the confidence in ~20 % of identified proteins. In total, 604 proteins were identified in nasal exosomes and the nasal exosomal proteome showed strong associations with immune-related functions, such as immune cell trafficking. Thirdly, a quantitative proteomics approach was used to determine alterations in the exosome proteome as a result of airway inflammatory disease. Serum-associated proteins and mucins were more abundant in the exosomes from subjects with respiratory diseases compared to healthy controls while proteins with antimicrobial functions and barrier-related proteins had decreased expression. Nasal exosomes were shown to induce the migration of innate immune cells, which may be important as the airway epithelium is the first line of defence against pathogens and allergens. The decreased expression in barrier and antimicrobial exosomal proteins in subjects with airway diseases, could possibly contribute to an increased susceptibility to infections, which have important clinical implications in disease progression. The online version of this article (doi:10.1186/s12967-016-0927-4) contains supplementary material, which is available to authorized users.