Limits...
Dual organism transcriptomics of airway epithelial cells interacting with conidia of Aspergillus fumigatus.

Oosthuizen JL, Gomez P, Ruan J, Hackett TL, Moore MM, Knight DA, Tebbutt SJ - PLoS ONE (2011)

Bottom Line: Concomitantly, A. fumigatus was found to up-regulate fungal pathways involved in iron acquisition, vacuolar acidification, and formate dehydrogenase activity.The up-regulation of IL-6 by epithelia and simultaneous activation of several pathways by fungal conidia warrants further investigation as we seek to better understand this interaction in both health and disease.The cellular response of the airway epithelium to A. fumigatus is important to understand if we are to improve host-pathogen outcomes.

View Article: PubMed Central - PubMed

Affiliation: UBC James Hogg Research Centre, Institute for HEART+LUNG Health, Providence Health Care, Vancouver, British Columbia, Canada.

ABSTRACT

Background: Given the complex nature of the responses that can occur in host-pathogen interactions, dual transcriptomics offers a powerful method of elucidating these interactions during infection. The gene expression patterns of Aspergillus fumigatus conidia or host cells have been reported in a number of previous studies, but each focused on only one of the interacting organisms. In the present study, we profiled simultaneously the transcriptional response of both A. fumigatus and human airway epithelial cells (AECs).

Methodology: 16HBE14o- transformed bronchial epithelial cells were incubated with A. fumigatus conidia at 37°C for 6 hours, followed by genome-wide transcriptome analysis using human and fungal microarrays. Differentially expressed gene lists were generated from the microarrays, from which biologically relevant themes were identified. Human and fungal candidate genes were selected for validation, using RT-qPCR, in both 16HBE14o- cells and primary AECs co-cultured with conidia.

Principal findings: We report that ontologies related to the innate immune response are activated by co-incubation with A. fumigatus condia, and interleukin-6 (IL-6) was confirmed to be up-regulated in primary AECs via RT-qPCR. Concomitantly, A. fumigatus was found to up-regulate fungal pathways involved in iron acquisition, vacuolar acidification, and formate dehydrogenase activity.

Conclusion: To our knowledge, this is the first study to apply a dual organism transcriptomics approach to interactions of A. fumigatus conidia and human airway epithelial cells. The up-regulation of IL-6 by epithelia and simultaneous activation of several pathways by fungal conidia warrants further investigation as we seek to better understand this interaction in both health and disease. The cellular response of the airway epithelium to A. fumigatus is important to understand if we are to improve host-pathogen outcomes.

Show MeSH

Related in: MedlinePlus

Localization of A. fumigatus conidia within the airway epithelial cell monolayer.GFP-expressing A. fumigatus conidia and primary AECs were co-incubated for 6 hours and treated with DAPI and monoclonal E-cadherin Alexa 594 antibody before visualization by confocal microscopy. Labeling of nuclei (blue) and the membrane tight junctional protein E-cadherin (red) allowed visualization of AECs. Some GFP-expressing A. fumigatus conidia (green) are found outside the cells, while others localize within the cell monolayer, in close association with AEC nuclei.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3105077&req=5

pone-0020527-g001: Localization of A. fumigatus conidia within the airway epithelial cell monolayer.GFP-expressing A. fumigatus conidia and primary AECs were co-incubated for 6 hours and treated with DAPI and monoclonal E-cadherin Alexa 594 antibody before visualization by confocal microscopy. Labeling of nuclei (blue) and the membrane tight junctional protein E-cadherin (red) allowed visualization of AECs. Some GFP-expressing A. fumigatus conidia (green) are found outside the cells, while others localize within the cell monolayer, in close association with AEC nuclei.

Mentions: To assess whether co-incubation would lead to the uptake of A. fumigatus conidia by primary human AECs, co-cultures were first visualized using incremental focal planes, to produce a Z-stack to form a three dimensional image by confocal microscopy. As shown in Figure 1, conidia expressing GFP co-incubated with AECs could be seen adjacent both to the cell plasma membrane identified by E-cadherin staining (red staining), and also to the AEC nucleus identified by DAPI (blue staining).


Dual organism transcriptomics of airway epithelial cells interacting with conidia of Aspergillus fumigatus.

Oosthuizen JL, Gomez P, Ruan J, Hackett TL, Moore MM, Knight DA, Tebbutt SJ - PLoS ONE (2011)

Localization of A. fumigatus conidia within the airway epithelial cell monolayer.GFP-expressing A. fumigatus conidia and primary AECs were co-incubated for 6 hours and treated with DAPI and monoclonal E-cadherin Alexa 594 antibody before visualization by confocal microscopy. Labeling of nuclei (blue) and the membrane tight junctional protein E-cadherin (red) allowed visualization of AECs. Some GFP-expressing A. fumigatus conidia (green) are found outside the cells, while others localize within the cell monolayer, in close association with AEC nuclei.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3105077&req=5

pone-0020527-g001: Localization of A. fumigatus conidia within the airway epithelial cell monolayer.GFP-expressing A. fumigatus conidia and primary AECs were co-incubated for 6 hours and treated with DAPI and monoclonal E-cadherin Alexa 594 antibody before visualization by confocal microscopy. Labeling of nuclei (blue) and the membrane tight junctional protein E-cadherin (red) allowed visualization of AECs. Some GFP-expressing A. fumigatus conidia (green) are found outside the cells, while others localize within the cell monolayer, in close association with AEC nuclei.
Mentions: To assess whether co-incubation would lead to the uptake of A. fumigatus conidia by primary human AECs, co-cultures were first visualized using incremental focal planes, to produce a Z-stack to form a three dimensional image by confocal microscopy. As shown in Figure 1, conidia expressing GFP co-incubated with AECs could be seen adjacent both to the cell plasma membrane identified by E-cadherin staining (red staining), and also to the AEC nucleus identified by DAPI (blue staining).

Bottom Line: Concomitantly, A. fumigatus was found to up-regulate fungal pathways involved in iron acquisition, vacuolar acidification, and formate dehydrogenase activity.The up-regulation of IL-6 by epithelia and simultaneous activation of several pathways by fungal conidia warrants further investigation as we seek to better understand this interaction in both health and disease.The cellular response of the airway epithelium to A. fumigatus is important to understand if we are to improve host-pathogen outcomes.

View Article: PubMed Central - PubMed

Affiliation: UBC James Hogg Research Centre, Institute for HEART+LUNG Health, Providence Health Care, Vancouver, British Columbia, Canada.

ABSTRACT

Background: Given the complex nature of the responses that can occur in host-pathogen interactions, dual transcriptomics offers a powerful method of elucidating these interactions during infection. The gene expression patterns of Aspergillus fumigatus conidia or host cells have been reported in a number of previous studies, but each focused on only one of the interacting organisms. In the present study, we profiled simultaneously the transcriptional response of both A. fumigatus and human airway epithelial cells (AECs).

Methodology: 16HBE14o- transformed bronchial epithelial cells were incubated with A. fumigatus conidia at 37°C for 6 hours, followed by genome-wide transcriptome analysis using human and fungal microarrays. Differentially expressed gene lists were generated from the microarrays, from which biologically relevant themes were identified. Human and fungal candidate genes were selected for validation, using RT-qPCR, in both 16HBE14o- cells and primary AECs co-cultured with conidia.

Principal findings: We report that ontologies related to the innate immune response are activated by co-incubation with A. fumigatus condia, and interleukin-6 (IL-6) was confirmed to be up-regulated in primary AECs via RT-qPCR. Concomitantly, A. fumigatus was found to up-regulate fungal pathways involved in iron acquisition, vacuolar acidification, and formate dehydrogenase activity.

Conclusion: To our knowledge, this is the first study to apply a dual organism transcriptomics approach to interactions of A. fumigatus conidia and human airway epithelial cells. The up-regulation of IL-6 by epithelia and simultaneous activation of several pathways by fungal conidia warrants further investigation as we seek to better understand this interaction in both health and disease. The cellular response of the airway epithelium to A. fumigatus is important to understand if we are to improve host-pathogen outcomes.

Show MeSH
Related in: MedlinePlus