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Functional genomics of Aspergillus fumigatus conidiospores interacting with human bronchial epithelial cells

ABSTRACT: The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. The transcriptional profiles of A. fumigatus conidiospores incubated in the presence or absence of human cells were compared, revealing significant changes in fungal gene expression in response to conidial interaction with cells.

ORGANISM(S): Aspergillus fumigatus

PROVIDER: GSE16630 | GEO | 2011/06/01

SECONDARY ACCESSION(S): PRJNA122795

REPOSITORIES: GEO

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Functional genomics of Aspergillus fumigatus conidiospores interacting with human bronchial epithelial cells

Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. The transcriptional profiles of A. fumigatus conidiospores incubated in the presence or absence of human cells were compared, revealing significant changes in fungal gene expression in response to conidial interaction with cells. Gene expression levels were investigated in A. fumigatus conidiospores incubated in the presence or absence of 16HBE cells for 6 hours. Four independent samples were analysed for each of these conditions, using JCVI PFGRC Aspergillus fumigatus Version 3 microarrays, with a dye-swap experimental design.

2011-06-01 | E-GEOD-16630 | biostudies-arrayexpress

Functional genomics of human bronchial epithelial cells interacting with conidiospores of Aspergillus fumigatus

2011-06-01 | GSE16628 | GEO

Functional genomics of human bronchial epithelial cells interacting with conidiospores of Aspergillus fumigatus

Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. Comparing the transcriptional profiles of control cells and cells exposed to A. fumigatus conidiospores using Agilent Whole Human Genome microarrays revealed significant changes in gene expression in response to the presence of conidiospores. The identification of biologically relevant responses validates this methodology, and motivates further work to characterize the interactions between A. fumigatus conidiospores and primary airway epithelial cells of normal and asthmatic origins. Gene expression levels were investigated in 16HBE cells incubated in the presence or absence of A. fumigatus conidiospores for 6 hours. Four independent cell samples were analysed for each of these conditions, using Agilent Whole Human Genome microarrays, with a one-colour experiemntal design.

2011-06-01 | E-GEOD-16628 | biostudies-arrayexpress

Functional genomics of human bronchial epithelial cells directly interacting with conidiospores of Aspergillus fumigatus

Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. Using fluorescence-activated cell sorting (FACS), cells directly interacting with conidiospores and cells not associated with any conidiospores were sorted into separate samples. Transcriptional profiling of these samples using Agilent Whole Human Genome microarrays revealed significant changes in gene expression in response to interaction with conidiospores. The identification of biologically relevant responses validates this methodology, and motivates further work to characterize the interactions between A. fumigatus conidiospores and primary airway epithelial cells of normal and asthmatic origins.

2010-06-04 | GSE16627 | GEO

Functional genomics of human bronchial epithelial cells directly interacting with conidiospores of Aspergillus fumigatus

Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. Using fluorescence-activated cell sorting (FACS), cells directly interacting with conidiospores and cells not associated with any conidiospores were sorted into separate samples. Transcriptional profiling of these samples using Agilent Whole Human Genome microarrays revealed significant changes in gene expression in response to interaction with conidiospores. The identification of biologically relevant responses validates this methodology, and motivates further work to characterize the interactions between A. fumigatus conidiospores and primary airway epithelial cells of normal and asthmatic origins. Four independent samples of 16HBE cells incubated with GFP-expressing Aspergillus fumigatus conidiospores for 6 hours were analyzed by flow cytometry and divided into negative and positive samples based on the presence of a GFP signal, representing cells with no direct contact with conidiospores and cells directly interacting with conidiospores, respectively. Gene expression was analyzed in these 8 samples using Agilent Whole Human Genome microarrays, with a one-colour experiemntal design.

2010-06-16 | E-GEOD-16627 | biostudies-arrayexpress

Transcriptional profiling of ATCC46645 strain and isogenic ΔpacC Aspergillus fumigatus mutant during murine infection

Project description:In lung diseases caused by the major mould pathogen Aspergillus fumigatus the pulmonary epithelium is destroyed by invasive growth of fungal hyphae, a process thought to require fungal proteases. Here we show that the A. fumigatus pH-responsive transcription factor PacC governs expression of secreted proteases during invasive lung infections and is required for epithelial invasion and pathogenicity. In addition, A. fumigatus ΔpacC mutants aberrantly remodel the fungal cell wall during infection. This study defines distinct PacC-mediated mechanisms of host damage during pulmonary aspergillosis. ch1: treatment protocol

2014-10-20 | GSE54810 | GEO

Transcriptional profiling of ATCC46645 strain and isogenic M-NM-^TpacC Aspergillus fumigatus mutant during murine infection

Project description:In lung diseases caused by the major mould pathogen Aspergillus fumigatus the pulmonary epithelium is destroyed by invasive growth of fungal hyphae, a process thought to require fungal proteases. Here we show that the A. fumigatus pH-responsive transcription factor PacC governs expression of secreted proteases during invasive lung infections and is required for epithelial invasion and pathogenicity. In addition, A. fumigatus M-NM-^TpacC mutants aberrantly remodel the fungal cell wall during infection. This study defines distinct PacC-mediated mechanisms of host damage during pulmonary aspergillosis. ch1: treatment protocol Temporal transcriptional profiling of ATCC46645 strain and isogenic M-NM-^TpacC Aspergillus fumigatus mutant during murine infection

2014-10-20 | E-GEOD-54810 | biostudies-arrayexpress

Transcription profiling of mouse C57Bl/6 or gp91phox-/- animals following exposure to conidia or polystyrene beads

Project description:The response of AM transcription to exposure to conidia is expected to provide information about the mechanism by which these cells prevent conidial germination and therefore invasive aspergillosis. Experiment Overall Design: Conidia from A. fumigatus or polystyrene beads were instilled into lungs of C57Bl/6 or gp91phox-/- mice to test the response of AM in a normal and immune compromised host to determine if different responses contributed to increased susceptibility of invasive aspergillosis. Following in vivo incubation for 0, 2, or 4 hours, AM RNA was collected and prepared for hybridization to mouse Affymetrix GeneChip arrays.

2008-06-17 | E-GEOD-8997 | biostudies-arrayexpress

RNA-Seq analysis of gene expression changes by wildtype and Nlrx1 deficient BEAS-2B airway epithelial cells in response to A. fumigatus viable conidia

Project description:Aspergillus fumigatus is an opportunistic fungal pathogen of the respiratory system that may cause invasive infection or an allergic response. Bronchial airway epithelial cells compromise the most abundant cell type of the pulmonary-air interface. Our recent findings suggest loss of Nlrx1 by BEAS-2B airway epithelial cells result in a hyper inflammatory response as well as decreased conidial processing. We challenged wildtype Nlrx1 deficient BEAS-2B cells with viable A. fumigatus AF293 conidia to identify early response differentially expressed genes and pathways between the two cell populations.

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Project description:Aspergillus fumigatus is the most important pulmonary fungal pathogen and is able to cause several diseases collectively called aspergillosis. Conidia is the most important infection structure making the initial contact with the human host. Here, we used a phylogenomic approach comparing proteins present in the A. fumigatus conidial surface, two closely related non-pathogenic species A. fischeri and A. oerlinghausenensis, and the far-related pathogenic A. lentulus. We were able to identify 62 proteins specifically expressed on A. fumigatus conidial surface. We deleted 42 of the encoding-genes and observed that many of them have altered susceptibility to macrophage killing, penetration and damage to epithelial cells, and cytokine production. We demonstrated that one of these genes encoding a glycosylasparaginase is modulating IL-1β levels and is important for the infection in an immunocompetent murine model. Our results provide opportunities for characterizing A. fumigatus effectors important for evasion and modulation of the immune response.

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