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Clinical

Gene expression profiling of differentiated HNECs stimulated by IL4, IL13, IFNalpha, IFNbeta, IFNgamma and controls - GSE19182

Purpose

In asthma, airway epithelium remodeling can already be detected during childhood, and epithelial cells are more susceptible to virus and oxidative stress. Their exact role in natural history and severity of children allergic respiratory disease remains however surprisingly unexplored. The aim of the study was to analyze dysfunctions of epithelium in dust mite allergic respiratory disease (rhinitis ± asthma) in children.

Hypothesis

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Experimental Design

Expression profilings of nasal epithelial cells collected by brushing were performed on Affymetrix Hugene 1.0 ST arrays. All allergic patients were sensitized to dust mite. 19 patients had an isolated allergic rhinitis (AR). 14 patients had AR associated with asthma. Patients were compared to 12 controls, their severity and control being assessed according to NAEPP and ARIA criteria. Infections by respiratory viruses were excluded by real-time PCR measurements. Each condition was performed in triplicates: total of 21 samples.

Experimental Variables

Presence of allergic rhinitis or asthma. Respiratory viruses infection excluded the participants.

Controls

12 primary cultures of human nasal epithelial cells stimulated with either interleukin (IL)-4, IL-13, interferon (IFN)-alpha, IFN-beta or IFN-gamma, or during in vitro differentiation.

Methods

Gene expression signatures of nasal brushings from children with dust mite-allergic rhinitis, associated or not associated with controlled or uncontrolled asthma. Supervised learning and unsupervised clustering were used to predict the different subgroups of patients and define altered signalling pathways.

Additional Information

This dataset is relevant to dust mite allergy and asthma.

Microarray
Affymetrix HuGene 1.0 ST v1
21 Samples Loaded: 21
Human (Homo sapiens)
human nasal epithelial cells
Allergy
Control HNECs and HNECs treated with 10 ng/mL IL4 or 10 ng/mL IL-13 were incubated for 24 h. HNECs treated with 1000 UI/mL IFNalpha, 1000 UI/mL IFNbeta, or 100 ng/mL IFNgamma were incubated for 6 h.
Absolutely RNA® Microprep Kit extraction of total RNA was performed according to the manufacturer's instructions
Label protocol: Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 300 ng total RNA. Hybridization protocol: Following fragmentation and labeling, 3ug of DNA were hybridized for 17 hr at 45C on Huge gene 1.0ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450. Scan protocol: GeneChips were scanned using the GeneChip Scanner 3000 7G (Affymetrix). Data processing: The data were analyzed with Expression Console using Affymetrix default analysis settings and Gene Level-RMA sketch as normalization method.
Sample Set Spreadsheet
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Samples Preview
Sample ID !Sample Title subject Cell type Sample Type Stimulation
GSM475535 01-D24-IL4 D24 HNECs Differentiated HNECs cultured in ALI between D15-D21 IL4
GSM475536 02-D24-IL13 D24 HNECs Differentiated HNECs cultured in ALI between D15-D21 IL13
GSM475537 03-D24-C D24 HNECs Differentiated HNECs cultured in ALI between D15-D21 C
GSM475538 04-D25-IL4 D25 HNECs Differentiated HNECs cultured in ALI between D15-D21 IL4
GSM475539 05-D25-IL13 D25 HNECs Differentiated HNECs cultured in ALI between D15-D21 IL13
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File
GSE19182 v1.csv
Sample Set Spreadsheet
GSE19182 v1.csv
Sample Set Spreadsheet
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Sample ID !Sample Title Subject Cell type Sample Type Stimulation
GSM475535
01-D24-IL4
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL4
GSM475536
02-D24-IL13
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL13
GSM475537
03-D24-C
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C
GSM475538
04-D25-IL4
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL4
GSM475539
05-D25-IL13
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL13
GSM475540
06-D25-C
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C
GSM475541
07-D26-IL4
D26
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL4
GSM475542
08-D26-IL13
D26
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IL13
GSM475543
09-D26-C
D26
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C
GSM475544
10-D21-IFNalpha
D21
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNalpha
GSM475545
11-D21-IFNbeta
D21
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNbeta
GSM475546
12-D21-IFNgamma
D21
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNgamma
GSM475547
13-D21-C
D21
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C
GSM475548
14-D24-IFNalpha
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNalpha
GSM475549
15-D24-IFNbeta
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNbeta
GSM475550
16-D24-IFNgamma
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNgamma
GSM475551
17-D24-C
D24
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C
GSM475552
18-D25-IFNalpha
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNalpha
GSM475553
19-D25-IFNbeta
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNbeta
GSM475554
20-D25-IFNgamma
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
IFNgamma
GSM475555
21-D25-C
D25
HNECs
Differentiated HNECs cultured in ALI between D15-D21
C

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