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Series GSE19392 Query DataSets for GSE19392
Status Public on Mar 30, 2010
Title Dynamic responses of primary human bronchial epithelial cells to influenza virus, viral RNA and interferon-beta
Organism Homo sapiens
Experiment type Expression profiling by array
Summary We defined the major transcriptional responses in primary human bronchial epithelial cells (HBECs) after either infection with influenza or treatment with relevant ligands. We used four different strategies, each highlighting distinct aspects of the response. (1) cells were infected with the wild-type PR8 influenza virus that can mount a complete replicative cycle. (2) cells were transfected with viral RNA (‘vRNA’) isolated from influenza particles. This does not result in the production of viral proteins or particles and identifies the effect of RNA-sensing pathways (e.g., RIG-I.). (3) Cells were treated with interferon beta (IFNb), to distinguish the portion of the response which is mediated through Type I IFNs. (4) Cells were infected with a PR8 virus lacking the NS1 gene (‘DNS1’). The NS1 protein normally inhibits vRNA- or IFNb-induced pathways, and its deletion can reveal an expanded response to infection.
 
Overall design HBECs were stimulated with a 15 minute pulse of 1000U/ml IFNß (PBL, Piscataway, NJ), 100ng/ml vRNA (purified directly from PR8 virus) with LTX transfection reagent (Invitrogen; Carlsbad, California), wild type H1N1 influenza (A/PR/8/34) or ?NS1 virus (PR8 with a deleted NS1 gene, gift from Dr. Garcia-Sastre). Viruses were used at a multiplicity of infection (moi) of 5. Control samples were incubated with media or LTX under the same conditions. Cells were washed, supplemented with warm media and harvested at 11 timepoints (0, .25, .5, 1, 1.5, 2, 4, 6, 8, 12, and 18 hours post-treatment). HBECs were seeded in 6 well plates at a concentration of 250,000/well 18 hours prior to stimultaion. Cells were stimulated with a 15 minute pulse of IFNb, vRNA, infected with PR8 influenza or NS1 deleted influenza, or mock treated
 
Contributor(s) Shapira S, Shum B, Hacohen N
Citation(s) 20064372
Submission date Dec 09, 2009
Last update date Jan 17, 2017
Contact name Nir Hacohen
E-mail(s) nhacohen@partners.org
Phone 617-724-3768
Organization name Broad Institute
Street address 7 Cambridge Center
City Cambridge
State/province MA
ZIP/Postal code 02129
Country USA
 
Platforms (1)
GPL3921 [HT_HG-U133A] Affymetrix HT Human Genome U133A Array
Samples (169)
GSM528620 HBEs treated with 100ul of pre trypsin media 100pre0.5_C11_1
GSM528621 HBEs treated with 100ul of pre trypsin media 100pre1.5_F12_1
GSM528622 HBEs treated with 100ul of pre trypsin media 100pre18_H4_1
Relations
BioProject PRJNA121751

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE19392_RAW.tar 374.4 Mb (http)(custom) TAR (of CEL)
Processed data included within Sample table
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