Comment[ArrayExpressAccession]	E-GEOD-43700
MAGE-TAB Version	1.1						
Public Release Date	2013-03-05
Investigation Title	Microarray analysis of IL-10 stimulated adherent peripheral blood mononuclear cells						
Comment[Submitted Name]	Microarray analysis of IL-10 stimulated adherent peripheral blood mononuclear cells
Experiment Description	The immune mechanisms that control resistance vs. susceptibility to mycobacterial infection in humans were investigated by studying leprosy skin lesions, the site where the battle between the host and the pathogen is joined.  Using an integrative genomics approach, we found an inverse correlation between of IFN-beta and IFN-gamma gene expression programs at the site of disease.  The Type II IFN, IFN-gamma and its downstream vitamin D-dependent antimicrobial genes were preferentially expressed in the lesions from patients with the self-healing tuberculoid form of the disease and mediated antimicrobial activity against the pathogen, Mycobacterium leprae in vitro.  In contrast, the Type I IFN, IFN-beta and its downstream genes, including IL-27 and IL-10, were induced in monocytes by M. leprae in vitro, and were preferentially expressed in the lesions of disseminated and progressive lepromatous form.  The IFN-gamma induced macrophage antimicrobial response was inhibited by IFN-beta/IL-10, by a mechanism involving blocking the generation of bioactive 1,25-dihyroxy vitamin D as well as inhibiting induction of antimicrobial peptides cathelicidin and DEFB4.  The ability of IFN-M-oM-^AM-" to inhibit the IFN-gamma induced vitamin D pathway including antimicrobial activity was reversed by neutralization of IL-10, suggesting a possible target for therapeutic intervention.  Finally, a common IFN-beta and IL-10 gene signature was identified in both the skin lesions of leprosy patients and in the peripheral blood of active tuberculosis patients.  Together these data suggest that the ability of IFN-beta to downregulate protective IFN-gamma responses provides one general mechanism by which some bacterial pathogens of humans evade protective host responses and contribute to pathogenesis. Peripheral blood monuclear cells derived through Ficoll-Hypaque from the blood of healthy human donors (n=4).  Cells adhered to tissue culture treated 6-well plates for 2 hours in 1% Fetal Bovine Serum (FBS) RPMI and then stimulated by IL-10 (R&D Systems) 10ng/ml or media alone in 10% FBS RPMI for 24 hours at 37M-BM-0C, 5% CO2.   After stimulation, cells harvested and monocytes isolated through CD14 positive selection (Miltenyi Biotec).  RNA from purified monocytes extracted by Trizol and purified by Qiagen RNeasy Kit.  RNA probe and microarray performed by UCLA Clinical Microarray Core using Ambion labeling kit and Affymetrix Human U133 Plus 2.0 array.						
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl					
Person Last Name	Montoya	Montoya	Teles				
Person First Name	Dennis	Dennis	Rosane				
Person Mid Initials			M				
Person Email	dmontoya@mednet.ucla.edu						
Person Affiliation	David Geffen School of Medicine at UCLA						
Person Phone	6269229221						
Person Address	David Geffen School of Medicine at UCLA, 615 Charles E Young Dr. S, Los Angeles, CA, USA						
Person Roles	submitter						
Protocol Name	P-GSE43700-1	P-GSE43700-6	P-GSE43700-5	P-GSE43700-2	P-GSE43700-3	P-GSE43700-4	P-GSE43700-7
Protocol Description	ID_REF =  VALUE = RMA normalized call = 	Performed by UCLA Clinical Microarray Core	Performed by UCLA Clinical Microarray Core on Affymetrix Human Genome U133 Plus 2.0 Array	Peripheral blood monuclear cells derived through Ficoll-Hypaque from the blood of healthy human donors (n=4). Cells adhered to tissue culture treated 6-well plates for 2 hours in 1% Fetal Bovine Serum (FBS) RPMI and then stimulated by IL-10 (R&D Systems) 10ng/ml or media alone in 10% FBS RPMI for 24 hours at 37M-0C, 5% CO2.	Trizol protocol for RNA isolation followed by purification of RNA by Qiagen Rneasy kit	Performed by UCLA Clinical Microarray Core using Ambion Labeling kit	Dchip analysis and RMA normalization using DNA-Chip Analyzer (Build date: Feb 21 2010, www.dchip.org)
Protocol Type	bioassay_data_transformation	image_aquisition	hybridization	specified_biomaterial_action	nucleic_acid_extraction	labeling	feature_extraction
Experimental Factor Name	STIMULATED WITH	ID					
Experimental Factor Type	stimulated with	id					
Publication Title	Type I Interferon Suppresses Type II Interferon-Triggered Human Anti-Mycobacterial Responses.						
Publication Author List	Teles RM, Graeber TG, Krutzik SR, Montoya D, Schenk M, Lee DJ, Komisopoulou E, Kelly-Scumpia K, Chun R, Iyer SS, Sarno EN, Rea TH, Hewison M, Adams JS, Popper SJ, Relman DA, Stenger S, Bloom BR, Cheng G, Modlin RL						
PubMed ID	23449998						
Publication DOI	10.1126/science.1233665						
Comment[SecondaryAccession]	GSE43700						
Comment[GEOReleaseDate]	2013-03-05						
Comment[ArrayExpressSubmissionDate]	2013-01-23						
Comment[GEOLastUpdateDate]	2013-03-07						
Comment[AEExperimentType]	transcription profiling by array						
SDRF File	E-GEOD-43700.sdrf.txt