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PLoS One. 2016 Apr 14;11(4):e0152865. doi: 10.1371/journal.pone.0152865. eCollection 2016.

Proteomic Characterization of Middle Ear Fluid Confirms Neutrophil Extracellular Traps as a Predominant Innate Immune Response in Chronic Otitis Media.

Author information

1
Sheikh Zayed Center for Pediatric Surgical Innovation, Children's National Health System, Washington, DC, United States of America.
2
Center for Genetic Medicine Research, Children's National Health System, Washington, DC, United States of America.
3
Division of Pediatric Otolaryngology, Children's National Health System, Washington, DC, United States of America.

Abstract

BACKGROUND:

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.

OBJECTIVE:

This study aimed at characterizing the proteome of MEEs from children with COM with the goal of elucidating important innate immune responses.

METHOD:

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.

RESULTS:

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.

CONCLUSION:

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.

PMID:
27078692
PMCID:
PMC4831838
DOI:
10.1371/journal.pone.0152865
[Indexed for MEDLINE]
Free PMC Article

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