SuperSILAC Quantitative Proteome Profiling of Murine Middle Ear Epithelial Cell Remodeling with NTHi
Document Type
Journal Article
Publication Date
2016
Journal
PLoS ONE
Volume
11
Issue
2
DOI
10.1371/journal.pone.0148612
Keywords
Haemophilus Infections--metabolism; Haemophilus Infections--pathology; Haemophilus influenzae--classification; Otitis Media with Effusion--metabolism; Otitis Media with Effusion--pathology; Proteome--metabolism
Abstract
Background
Chronic Otitis Media with effusion (COME) develops after sustained inflammation and is characterized by secretory middle ear epithelial metaplasia and effusion, most frequently mucoid. Non-typeable Haemophilus influenzae (NTHi), the most common acute Otitis Media (OM) pathogen, is postulated to promote middle ear epithelial remodeling in the progression of OM from acute to chronic. The goals of this study were to examine histopathological and quantitative proteomic epithelial effects of NTHi challenge in a murine middle ear epithelial cell line.
Methods
NTHi lysates were generated and used to stimulate murine epithelial cells (mMEEC) cultured at air-liquid interface over 48 hours– 1 week. Conditional quantitative Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) of cell lysates was performed to interrogate the global protein production in the cells, using the SuperSILAC technique. Histology of the epithelium over time was done to measure bacterial dependent remodeling.
Results
Mass spectrometry analysis identified 2,565 proteins across samples, of which 74 exhibited differential enrichment or depletion in cell lysates (+/-2.0 fold-change; p value
Conclusions
NTHi lysates drive pathways responsible of cell remodeling in murine middle ear epithelium which likely contributes to observed epithelial hyperplasia in vitro. Further elucidation of these mediators will be critical in understanding the progression of OM from acute to chronic at the molecular level.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
APA Citation
Val, S., Burgett, K., Brown, K. J., & Preciado, D. (2016). SuperSILAC Quantitative Proteome Profiling of Murine Middle Ear Epithelial Cell Remodeling with NTHi. PLoS ONE, 11(2), e0148612. http://doi.org/10.1371/journal.pone.0148612
Peer Reviewed
1
Open Access
1
Comments
Reproduced with permission of PLoS ONE.