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Interaction with Human Bronchial Epithelial Cells in vitro Affects Gene Expression in Moraxella catarrhalis

ABSTRACT: In this study, DNA microarray technology was used to measure global gene expression in M. catarrhalis cells that had attached to human bronchial epithelial cells in culture.

ORGANISM(S): Moraxella catarrhalis

PROVIDER: GSE12990 | GEO | 2014/03/24

SECONDARY ACCESSION(S): PRJNA112599

REPOSITORIES: GEO

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Interaction with Human Bronchial Epithelial Cells in vitro Affects Gene Expression in Moraxella catarrhalis

Project description:In this study, DNA microarray technology was used to measure global gene expression in M. catarrhalis cells that had attached to human bronchial epithelial cells in culture. M. catarrhalis ATCC 43617 was allowed to attach (MOI~30) to a monolayer of 16HBE14 human bronchial epithelial cells for 2 hr in OmniTray cell culture plates. Additional portions of these bacteria were incubated in tissue culture medium in 75-cm2 cell culture flasks in the absence of 16HBE14 cells for 2 hr. After this incubation period, the medium in the flask with the 16HBE14 monolayer was decanted and replaced with tissue culture medium containing 25 mM sodium azide (to protect RNA). The 16HBE14 monolayer and attached bacteria were released from the plastic by scraping. After low-speed centrifugation to collect these bronchial cells and attached bacteria, the cell pellet was re-suspended in TE containing 1% saponin (which lyses the human cells). After a 10-min incubation at 37M-BM-0C, the bacterial cells were collected by centrifugation. The bacteria growing in the flask without human cells were treated identically and then harvested by centrifugation. Total RNA was extracted from both sets of bacterial cells by using the Qiagen RNeasy Midi kit. DNA microarray analysis, slide scanning, and statistical analysis of the data were performed. RNA was isolated four times and used in five DNA microarray hybridizations; M-bM-^@M-^\dye swapM-bM-^@M-^] experiments were performed twice. Two additional independent experiments were performed to isolate two sets of RNA samples for qRT-PCR analysis to validate DNA microarray data.

2014-03-24 | E-GEOD-12990 | biostudies-arrayexpress

Moraxella catarrhalis

Project description:Interaction with Human Bronchial Epithelial Cells in vitro Affects Gene Expression in Moraxella catarrhalis

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Deciphering the genetic basis of Moraxella catarrhalis complement resistance: Gene expression analysis of Moraxella catarrhalis BBH18 upon exposure to normal human serum.

Project description:The complement system is an important part of the innate defense against invading pathogens. The ability to resist complement-mediated killing is considered to be an important virulence trait for the human-restricted respiratory tract pathogen M. catarrhalis, as most disease-associated M. catarrhalis isolates are complement-resistant. Here we studied the molecular basis of M. catarrhalis complement-resistance by transcriptome profiling upon exposure to 10% normal human serum (NHS).

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Project description:This series of microarrays explores the genes regulated by the Zur gene of Moraxella catarrhalis.

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Expression profiling of Moraxella catarrhalis RH4 during in vitro growth in brain heart infusion (BHI) medium

Project description:Transcriptome analysis of M. catarrhalis RH4 predicted coding sequences and clustered regularly interspaced short palindromic repeats (CRISPRs) regions. The results described in this study are further discussed in De Vries, S.P.W., van Hijum, S.A.F.T., Schueler, W., Riesbeck, K., Hays, J.P., Hermans, P.W.M., Bootsma, H.J.; Genome analysis of Moraxella catarrhalis strain RH4: a human respiratory tract pathogen; Journal of Bacteriology

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Gene Expression Profile of Moraxella catarrhalis Grown in Pooled Human Sputum

Project description:Comparison of the gene expression profile of Moraxella catarrhalis grown in the presence of 20% pooled human sputum in chemically-defined medium relative to Moraxella catarrhalis grown in chemically-defined medium alone.

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