ArticleInternational journal of pediatric otorhinolaryngology2016
A proteomic characterization of NTHi lysates.
Article in International journal of pediatric otorhinolaryngology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 5 citations in OpenAlex.
- Review
- Learning from -omics strategies applied to uncoverComputational and structural biotechnology journal · 2021Review
- MUC5B induces in vitro neutrophil extracellular trap formation: Implication in otitis media.Laryngoscope investigative otolaryngology · 2020Article
- Nontypeable Haemophilus influenzae lysates increase heterogeneous nuclear ribonucleoprotein secretion and exosome release in human middle-ear epithelial cells.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2018Article
- Purification and characterization of microRNAs within middle ear fluid exosomes: implication in otitis media pathophysiology.Pediatric research · 2017Article
- SuperSILAC Quantitative Proteome Profiling of Murine Middle Ear Epithelial Cell Remodeling with NTHi.PloS one · 2016Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
backgroundNon-typeable Haemophilus influenzae (NTHi) is a ubiquitous bacterial pathogen which accounts for a majority of human upper respiratory tract infections. Laboratory lysate preparations from this bacterium are commonly utilized to investigate the promulgation of inflammatory responses in respiratory and middle ear epithelium both in vivo and in vitro. We undertook an unbiased proteomics based analysis of NTHi lysate preps to: (a) identify abundant bacterial proteins present in these lysates that could play a role in NTHi biological effects and (b) determine the protein content variability in different lysate prep batches from the same NTHI strain. STUDY
designProteomic analysis of laboratory NTHi lysate preparations from clinical strain 12.
methodsNTHi lysates were denatured, gel-fractionated, digested by trypsin and the generated peptides were identified using a liquid chromatography tandem mass spectrometry (LC-MS/MS). Western blot analyses for the important proinflammatory enhancer, outer membrane protein 6 (OMP6), was performed to validate the MS findings. Luciferase assays for NF-kB activation were used to measure the pro-inflammatory biologic effects from each NTHi lysate preparation.
resultsThe MS identified 793 unique NTHi proteins. Most common and abundant proteins found have been described to either contribute to biofilm formation, elude the innate immune system, or activate epithelial pro-inflammatory pathways such as Toll Like Receptor 2 (TLR-2) signaling and NF-kB transcription factor. Strong positive signal for OMP6 was found in each of the NTHi lysate preparations. Significant NF-kB promoter response activation as expected with NTHi stimulation over control was also noted for each NTHi lysate preparation.
conclusionsProteomics was a successful technique to broadly define the protein content of NTHi lysates. This is the first report of the proteome of NTHi lysates widely used in laboratories to study the biological effect of NTHi. Despite the variability of the protein composition from different preps, all the batches of NTHi lysates induced similar NFκB activation. LEVEL OF EVIDENCE: NA.
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