 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Aug 03, 2018 |
| Title |
Gene Expression of Primary Human Type I Alveolar Epithelial Cells Exposed to Bacillus anthracis, Sterne endospores |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
The lung is the entry site for Bacillus anthracis in inhalation anthrax, the most deadly form of the disease. Spores must escape through the alveolar epithelial cell (AEC) barrier and migrate to regional lymph nodes, germinate and enter the circulatory system to cause disease. Several mechanisms to explain alveolar escape have been postulated, and all these tacitly involve the AEC barrier. In this study, we incorporate our primary human type I AEC model, microarray gene profiling and gene enrichment analysis to study the response of AEC to B. anthracis, (Sterne) spores at 4 and 24 hours post-exposure. Spore exposure altered gene expression in AEC after 4 and 24 hours and differentially expressed genes (±1.3 fold, p ≤ 0.05) included CCL4/MIP-1β (4 hours), CXCL8/IL-8 (4 and 24 hours) and CXCL5/ENA-78 (24 hours). Gene enrichment analysis revealed that pathways involving cytokine or chemokine activity, receptor binding, and innate immune responses to infection were prominent. Microarray results were confirmed by qRT-PCR and multiplex ELISA assays. Chemotaxis assays demonstrated that spores induced the release of biologically active neutrophil and monocyte chemokines, and that CXCL8/IL-8 was the major neutrophil chemokine. The small or sub-chemotactic doses of CXCL5/ENA-78, CXCL3/GROββ and CCL20/MIP-3α may contribute to chemotaxis by priming effects. These data provide the first whole transcriptomic description of the human type I AEC initial response to B. anthracis spore exposure, and contribute to an increased understanding of the role of AEC in the pathogenesis of inhalational anthrax.
We used microarrays to create a whole transcriptomic description of the response of primary human type I alveolar epithelial cells to B. anthracis spore exposure and demonstrated that several of the most upregulated differentially expressed genes included those for neutrophil and monocyte chemokines.
|
| |
|
| Overall design |
Primary human type I alveolar epithelial cells isolated from the lungs of deceased donors were infected with 1 × 10^6 cfu/ml (MOI ≈ 2) Bacillus anthracis, Sterne strain 7702 (pX01+, pX02-) endospores in fresh growth medium containing antibiotics, and incubated for 4 and 24 h in a humidified incubator at 37°C in 5% CO2. Mock treated (negative control) cells received a volume of spore diluent (sterile di-water) equal to that of the spore treated cells. At the end of incubation, the cell monolayers were washed twice with PBS and lysed by addition of 700 μl/well QIAzol lysis reagent. Samples were processed into purified total RNA, which was reverse transcribed into cDNA and further amplified. Samples were fragmented and biotinylated for hybridization with Affymetrix GeneChip HuGene 2.0 ST Array, Format 100, chips. Array data was log2 transformed, quantile normalized and had batch effects removed for final analysis.
|
| |
|
| Contributor(s) |
Booth JL, Duggan ES, Metcalf JP, Burian DM, Dozmorov MG |
| Citation(s) |
29704667 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| U19 AI062629 |
Mechanisms by which B. anthracis Spores Escape the Lung |
University of Oklahoma Health Sciences Center |
JORDAN PATRICK METCALF |
|
| |
| Submission date |
Aug 01, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
J. Leland Booth |
| E-mail(s) |
john-booth@ouhsc.edu
|
| Phone |
405-271-1966
|
| Organization name |
Oklahoma University Health Sciences Center
|
| Department |
Internal Medicine Dept., Pulmonary and Critical Care Div.
|
| Lab |
JPMetcalf
|
| Street address |
800 N. Research Parkway
|
| City |
Oklahoma City |
| State/province |
OK |
| ZIP/Postal code |
73104 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL16686 |
[HuGene-2_0-st] Affymetrix Human Gene 2.0 ST Array [transcript (gene) version] |
|
| Samples (12)
|
| GSM2723967 |
AEC donor 1, +Spores, 4hr |
| GSM2723968 |
AEC donor 2, +Spores, 4hr |
| GSM2723969 |
AEC donor 3, +Spores, 4hr |
| GSM2723970 |
AEC donor 1, Mock, 24hr |
| GSM2723971 |
AEC donor 2, Mock, 24hr |
| GSM2723972 |
AEC donor 3, Mock, 24hr |
| GSM2723973 |
AEC donor 1, +Spores, 24hr |
| GSM2723974 |
AEC donor 2, +Spores, 24hr |
| GSM2723975 |
AEC donor 3, +Spores, 24hr |
|
| Relations |
| BioProject |
PRJNA396684 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE102106_RAW.tar |
89.9 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
|
|
|
|
 |
Less...
More...