A proteomic characterization of NTHi lysates.

摘要: Abstract Background Non-typeable Haemophilus influenzae (NTHi) is a ubiquitous bacterial pathogen which accounts for majority of human upper respiratory tract infections. Laboratory lysate preparations from this bacterium are commonly utilized to investigate the promulgation inflammatory responses in and middle ear epithelium both vivo vitro. We undertook an unbiased proteomics based analysis NTHi preps to: (a) identify abundant proteins present these lysates that could play role biological effects (b) determine protein content variability different prep batches same NTHI strain. Study design Proteomic laboratory clinical strain 12. Methods were denatured, gel-fractionated, digested by trypsin generated peptides identified using liquid chromatography tandem mass spectrometry (LC–MS/MS). Western blot analyses important proinflammatory enhancer, outer membrane 6 (OMP6), was performed validate MS findings. Luciferase assays NF-kB activation used measure pro-inflammatory biologic each preparation. Results The 793 unique proteins. Most common found have been described either contribute biofilm formation, elude innate immune system, or activate epithelial pathways such as Toll Like Receptor 2 (TLR-2) signaling transcription factor. Strong positive signal OMP6 preparations. Significant promoter response expected with stimulation over control also noted Conclusions Proteomics successful technique broadly define lysates. This first report proteome widely laboratories study effect NTHi. Despite composition preps, all induced similar NFκB activation. Level evidence NA