 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Nov 16, 2009 |
| Title |
Comparative Analysis of Genes Induced by Respiratory Syncytial Virus and DsRNA in Human Epithelial Cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
Epithelial cells are the primary target of respiratory viral infections and play a pivotal role in virus-induced lung inflammation and in anti viral immune response. A common signal for the presence of viral infections and induction of inflammation is recognition of double stranded RNA (dsRNA). Thus far, there has not been a high-throughput transcrptome analysis of RSV- or dsRNA-induced genes in primary human bronchial epithelial cells (PHBE), nor there has been a comparison between dsRNA- and RSV-induced genes. To establish the transcriptome profiles and to determine the contribution of dsRNA in the induction of inflammation during respiratory virus infection, we compared the gene expression profiles of PHBE cells that were infected with RSV or were treated with dsRNA. Our transcriptome analysis showed that RSV infection and and dsRNA treatment induced up-regulation of 2024 and 159 genes in PHBE respectively. Comparison of genes revealed that RSV and dsRNA commonly induced 80 genes in PHBE cells. The common up-regulated genes were functionally grouped in multiple response pathways involved in inflammation and immune responses. Interestingly, there were several previously unreported genes that were up-regulated in primary human epithelial cells that are relevant to a TH2 allergic phenotype. This comparison of a high-throughput gene expression study offers a comprehensive view of transcriptional changes induced by dsRNA and RSV, and importantly compares dsRNA-induced genes with RSV-induced genes in PHBE cells.
|
| |
|
| Overall design |
Primary human bronchial epithelial cells were left untreated, were treated with 1 ug/ml dsRNA (poly I:C), or were infected with RSV at multiplicity of infection (MOI) of 2.5 plaque forming unit (pfu)/cell. After 2 hr for dsRNA and 24 hr for RSV, total cellular RNA was isolated using the TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturers’ instructions.
Gene expression analysis was conducted using Agilent Human1Av2 arrays two color system (Agilent Technologies, Palo Alto, CA). Dye swap were performed to eliminate dye bias. Total RNA was amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 500ng of total RNA, Cy3 or Cy5 labeled cRNA was produced according to manufacturer’s protocol. For each two color comparison, 750ng of each Cy3 and Cy5 labeled cRNAs were mixed and fragmented using the Agilent In Situ Hybridization Kit protocol. Hybridizations were performed for 17 hours in a rotating hybridization oven using the Agilent 60-mer oligo microarray processing protocol. Slides were washed as indicated in this protocol and then scanned with an Agilent Scanner. Data was obtained using the Agilent Feature Extraction software (v7.5 and 9.1), using defaults for all parameters.
|
| |
|
| Contributor(s) |
Limmon GV, McCann KL, Andrews DM, Imani F |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Jul 17, 2008 |
| Last update date |
Dec 06, 2012 |
| Contact name |
NIEHS Microarray Core |
| E-mail(s) |
microarray@niehs.nih.gov, liuliw@niehs.nih.gov
|
| Organization name |
NIEHS
|
| Department |
DIR
|
| Lab |
Microarray Core
|
| Street address |
111 T.W. Alexander Drive
|
| City |
RTP |
| State/province |
NC |
| ZIP/Postal code |
27709 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL887 |
Agilent-012097 Human 1A Microarray (V2) G4110B (Feature Number version) |
|
| Samples (4)
|
|
| Relations |
| BioProject |
PRJNA113755 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE12144_RAW.tar |
127.8 Mb |
(http)(custom) |
TAR (of TIFF, TXT) |
| Processed data included within Sample table |
|
|
|
|
 |
Less...
More...