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Exosomes in the nose induce immune cell trafficking and harbour an altered protein cargo in chronic airway inflammation.

Journal article
Authors Cecilia Lässer
Serena O'Neil
Ganesh V Shelke
Carina Sihlbom
Sara Folkesson Hansson
Yong Song Gho
Bo Lundbäck
Jan Lötvall
Published in Journal of Translational Medicine
Volume 14
Issue 1
Pages 181
ISSN 1479-5876
Publication year 2016
Published at Krefting Research Centre
Core Facilities, Proteomics
Pages 181
Language en
Links dx.doi.org/10.1186/s12967-016-0927-...
Keywords Asthma, Chronic rhinosinusitis, Exclusion list, Exosomes, Extracellular vesicles, Cell migration, Mass spectrometry, Nasal lavage fluid, Proteomics, Tandem mass tags
Subject categories Cell Biology, Respiratory Medicine and Allergy

Abstract

BACKGROUND: 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)]. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

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