Comment[ArrayExpressAccession]	E-GEOD-19772
Investigation Title	Expression Data From HCMV-Infected Human Monocytes 48 Hours Post-Infection: Role of PI(3)K
Comment[Submitted Name]	Expression Data From HCMV-Infected Human Monocytes 48 Hours Post-Infection: Role of PI(3)K
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Experimental Design	transcription profiling by array
Experimental Design Term Source REF	EFO
Public Release Date	2010-01-06
Experiment Description	Human cytomegalovirus (HCMV) induces pro-inflammatory monocytes following infection and we have evidence that phosphatidylinositol 3-kinase [PI(3)K] is a key mediator in this activation. To begin to address how this signalling pathway is responsible for the functional changes in infected monocytes, we examined the role this pathway played in the transcriptome of infected monocytes. Global transcriptional profiling using cDNA microarrays revealed a significant number of genes were regulated in a PI(3)K-dependent manner, identifying this pathway as a key cellular control point in the conversion of monocytes to an activated pro-inflammatory state following HCMV infection.  Keywords: Disease state To begin to globally define how PI(3)K is involved in the HCMV-induced changes in monocyte function, we performed a transcriptome analysis in the presence of an inhibitor to the PI(3)K signalling pathway. Specifically, a cDNA microarray containing 12,626 unique probe sets was utilized to assess the modulation of the monocyte transcriptome at 48 hours post-infection in the presence of the pharmacological agent LY294002 (LY), a PI(3)K inhibitor. A total of 2 replicates from HCMV-infected monocytes, 2 replicates from LY-pretreated infected monocytes, and 2 replicates from mock-infected monocytes were analyzed in this study.							
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Person Last Name	Yurochko	Bivins-Smith	Smith	Yurochko				
Person First Name	Andrew	Elizabeth	M	Andrew				
Person Mid Initials	D.	R	S	D				
Person Email	ayuroc@lsuhsc.edu							
Person Affiliation	LSUHSC-S							
Person Phone	318-675-8332							
Person Fax	318-675-5764							
Person Address	Microbiology & Immunology, LSUHSC-S, 1501 Kings Highway, Shreveport, LA, USA							
Person Roles	submitter							
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Protocol Name	P-GSE19772-1	P-GSE19772-6	P-GSE19772-3	P-GSE19772-8	P-GSE19772-7	P-GSE19772-2	P-GSE19772-4	P-GSE19772-5
Protocol Description	ID_REF = <br>VALUE = MAS5.0 signal intensity<br>ABS_CALL = the call in an absolute analysis that indicates if the transcript was present (P), absent (A), or marginal (M)<br>DETECTION P-VALUE = p-value that indicates the significance level of the detection call	Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45C on the GeneChip Human U95Av2 Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.	Monocytes were incubated in RPMI supplemented with 10% human AB serum at 37C and 5% CO2.	The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling with a target intensity value of 2500 as normalization method.	GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.	Isolated monocytes were pretreated with or without LY294002. Cells wre HCMV infected (MOI 15) or mock infected, and incubated adherently at 37C for 48 hours.	RNA STAT-60 extraction of total RNA was performed according to the manufacturer's instructions.	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
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Protocol Type	bioassay_data_transformation	hybridization	grow	feature_extraction	image_aquisition	specified_biomaterial_action	nucleic_acid_extraction	labeling
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Experimental Factor Name	PRETREATMENT	INFECTION						
Experimental Factor Type	pretreatment	infection						
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Comment[SecondaryAccession]	GSE19772							
Comment[GEOLastUpdateDate]	2010-01-06
Comment[GEOReleaseDate]	2010-01-07
Comment[ArrayExpressSubmissionDate]	2010-01-06
SDRF File	E-GEOD-19772.sdrf.txt