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Expression data from Middle Ear Mucosal Metaplasia in Mice

ABSTRACT: Chronic Otitis Media (OM) 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 OM pathogen, is known to activate inflammation and mucin expression in vitro and in animal models of OM. The goals of this study were to: examine expression profiling epithelial effects of NTHi challenge in murine middle ears. We used microarrays to detail examine the global programme of gene expression underlying epithelial effects of NTHi challenge in murine middle ears during this study.

ORGANISM(S): Mus musculus

PROVIDER: GSE40087 | GEO | 2012/11/13

SECONDARY ACCESSION(S): PRJNA172758

REPOSITORIES: GEO

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Expression data from Middle Ear Mucosal Metaplasia in Mice

Project description:Chronic Otitis Media (OM) 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 OM pathogen, is known to activate inflammation and mucin expression in vitro and in animal models of OM. The goals of this study were to: examine expression profiling epithelial effects of NTHi challenge in murine middle ears. We used microarrays to detail examine the global programme of gene expression underlying epithelial effects of NTHi challenge in murine middle ears during this study. Weekly transtympanic inoculation of Balb/c mice with 300 Âµg/ml of NTHi lysates vs saline was performed. Bacteria were grown on chocolate agar at 37ÂºC in 5% CO2 overnight and inoculated in brain heart infusion (BHI) broth supplemented with 3.5 mg of nicotinamide adenine dinucleotide per ml. After overnight incubation, bacteria were subcultured into 5 ml of fresh brain heart infusion (BHI) and upon reaching log phase growth, NTHi were washed and suspended in phosphate-buffered saline (PBS) followed by sonication for lysis. Three transtympanic inoculation of 6 Balb/c mice middle ears (3 animals, 6 ears) with 50 uL of 300 ug/ml of NTHi bacterial lysate and 6 Balb/c mice middle ears (3 animals, 6 ears) with 50 uL of 1X phosphate buffered saline (PBS) were carried out weekly over 4 weeks (injection on days 7, 14, and 21). On day 28, the mice were euthanized and their bullae harvested. Expression microarray analysis was performed at 1 and 7 days. Microarray findings were validated in independent animal samples and in a cultured murine middle ear epithelial cell (mMEEC) line.

2012-11-13 | E-GEOD-40087 | biostudies-arrayexpress

A biphasic epigenetic switch controls immunoevasion, virulence and niche adapatation in non-typeable Haemophilus influenzae

Project description:Non-typeable Haemophilus influenzae (NTHi) contains an N6-adenine DNA-methyltransferase (ModA), that is subject to phase variable expression (random ON/OFF switching). Five modA alleles, modA2, 4, 5, 9 and 10, account for over two-thirds of clinical otitis media isolates surveyed. Single Molecule Real Time (SMRT) methylome analysis identified the DNA recognition motifs for all five of these modA alleles. Phase variation of these alleles regulated multiple proteins, including vaccine candidates. ON/OFF switching of modA alleles resulted in differential regulation of key virulence phenotypes, such as antibiotic resistance (modA2, 5, 10), biofilm formation (modA2) and immunoevasion (modA4). Analysis of the modA2 strain, 723, in the chinchilla model of otitis media showed a clear selection for switching from modA2OFF to ON in the middle ear. This is the first report of a biphasic epigenetic switch controlling bacterial virulence, immunoevasion and niche adaptation in an animal model system Direct comparison of biological triplicates of wild type and mutant strains

2015-06-12 | E-GEOD-69831 | biostudies-arrayexpress

A biphasic epigenetic switch controls immunoevasion, virulence and niche adapatation in non-typeable Haemophilus influenzae

2015-06-12 | GSE69831 | GEO

Mouse Middle Ear Infection with NTHi

Project description:NTHi bacteria or saline were inoculated into the middle ears of mice. Mice were sacrificed at various times to monitor the course of infection. We used Mouse430_2 arrays to profile gene expression in response to bacterial infection

2015-09-01 | GSE66141 | GEO

Influenza-induced inflammation drives pneumococcal otitis media

Project description:Influenza A virus (IAV) predisposes individuals to secondary infections with the bacterium Streptococcus pneumoniae (the pneumococcus). Infections may manifest as pneumonia, sepsis, meningitis or otitis media (OM). It remains controversial as to whether secondary pneumococcal disease is due to the induction of an aberrant immune response or IAV induced immunosuppression. Moreover, as the majority of studies have been performed in the context of pneumococcal pneumonia, it remains unclear how far these findings can be extrapolated to other pneumococcal disease phenotypes. Here, we demonstrate that the viral hemagglutinin (HA) mediates bacterial OM by inducing a pro-inflammatory response in the middle ear cavity in a replication-dependent manner. Importantly, our findings show that it is the inflammatory response that mediates pneumococcal replication; not viral suppression of the immune system or epithelial damage. This study provide the first evidence that HA induced inflammation drives pneumococcal replication in the middle ear cavity, which has important consequences to the treatment of pneumococcal OM.

2013-01-01 | GSE41098 | GEO

Mouse Middle Ear Infection with NTHi

2015-09-01 | E-GEOD-66141 | biostudies-arrayexpress

The transcriptional landscape of the murine middle ear epithelium in vitro

Project description:Middle ear epithelium (MEE) is an extended part of the respiratory mucosa and mucociliary differentiation of MEE is crucial for the maintenance of homeostasis and sterility of the middle ear cavity. Previously we developed an in vitro model of murine MEE using air-liquid interface (ALI) culture that replicates aspects of the in vivo middle ear. Defects in MEE function can lead to the development of otitis media (OM). Decoding differential gene expression (DGE) across the MEE during mucociliary differentiation is valuable in understanding the mechanism underpinning the development of OM. We used ClariomTM S Mouse assay to understand the global gene expression at different time points of murine MEE differentiation at ALI culture and to identify sets of genes up-regulated and down-regulated during this process.

2021-04-20 | GSE150764 | GEO

Identification of a novel haem-binding protein produced by Haemophilus haemolyticus

Project description:Characterization of a novel haem-binding protein produced by Haemophilus haemolyticus, isolated and purified from non-typeable Haemophilus influenzae (NTHi)-inhibitory substance.

2023-04-27 | PXD013342 | Pride

Beta- lactam antibiotics stimulate biofilm formation in non-typeable Haemophilus influenzae by up-regulating carbohydrate metabolism

Project description:Non-typeable Haemophilus influenzae (NTHi) is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth) stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10M-BM-5g/mL) of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended. 12 samples

2014-06-04 | E-GEOD-58213 | biostudies-arrayexpress

Identification of a novel haem-binding protein produced by Haemophilus haemolyticus

Project description:Characterization of a novel haem-binding protein that is secreted by two Haemophilus haemolyticus isolates, BW1 and RHH122, and that inhibits the growth of the pathogen non-typeable Haemophilus influenzae (NTHi) in vitro.

2019-05-29 | PXD013687 | Pride

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