Comment[ArrayExpressAccession]	E-GEOD-70146
MAGE-TAB Version	1.1					
Public Release Date	2015-12-31
Investigation Title	Difference of gene expression between LPS-stimulated monocytes cultured in the presence NRBCs and those cultured in the absence of NRBCs					
Comment[Submitted Name]	Difference of gene expression between LPS-stimulated monocytes cultured in the presence NRBCs and those cultured in the absence of NRBCs
Experiment Description	We found that human cord blood nucleated red blood cells (NRBCs) have a regulatory function in the innate immune reaction. NRBCs suppressed the production of inflammatory cytokines including TNF-a and IL-1b from monocytes in response to lipopolysaccharide (LPS). NRBCs exerted the regulatory function even without cell-to-cell contact with monocytes. On the other hand, IL-10 production from monocytes by the stimulation of LPS in the presence of NRBCs was higher than that of LPS-stimulated monocytes cultured in the absence of NRBCs. Addition of an anti-IL-10 receptor blocking antibody restored the inflammatory cytokine production from monocytes, suggesting that the functional change of monocytes caused by the interaction of NRBCs was characterized and mediated by the increased IL-10 production. Whole-genome microarray analysis revealed that monocytes expressed increased amounts of IL-10 superfamily genes after the interaction with NRBCs. IL-19, one of the IL-10 superfamily, enhanced IL-10 production from monocytes, which suggested cooperative role of IL-10 superfamily in the suppression of inflammatory cytokine production from monocytes. Arginase which was reported to play an important role in the suppressive function of NRBCs on monocytes in mice was found to play no significant role in human monocytes. NRBCs seem to have a regulatory role to suppress vigorous innate immune reaction which can be harmful to the fetuses via some unknown soluble factor which enhances the production of IL-10 and IL-10 family cytokines in monocytes. Human LPS-stimulated monocytes were collected after culture in a condition without and with NRBCs in transwell culture system. Only one experiment was performed.					
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	Li	Cui	Takada			
Person First Name	Cui	Lili	Hidetoshi			
Person Mid Initials	Li					
Person Email	geo@ncbi.nlm.nih.gov					
Person Affiliation	Kyushu university					
Person Address	Pediatrics, Kyushu university, Maidashi3-1-1, Fukuoka, Japan					
Person Roles	submitter					
Protocol Name	P-GSE70146-1	P-GSE70146-4	P-GSE70146-5	P-GSE70146-2	P-GSE70146-3	P-GSE70146-6
Protocol Description	The scanned images were analyzed with Feature Extraction Software 9.5.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. The raw signal intensities and flags for each probe were calculated from the hybridization intensities and spot information according to the procedures recommended by Agilent Technologies using the Flag criteria in the GeneSpring Software. In addition, the raw signal intensities of two samples were log2-transformed and normalized by the quantile algorithm with the Bioconductor. ID_REF =  VALUE = quantile normalized signal (non-log scaled) and ABS CALL. ABS_CALL = 	cRNA was amplified and labelled using a Low input Quick Amp Labelling Kit (Agilent Technologies).	cRNA was hybridized to a 60K 60-mer oligomicroarray (SurePrint G3 Human Gene Expression Microarray 8x60K v2; Agilent Technologies) according to the manufacturer's instructions.	Fresh human monocytes were stimulated with LPS (100ng/ml) alone or transwell cocultured with NRBCs, which derived from cord blood for 36h. Monocytes were collected after culture.	Total RNA was isolated from cells using TRIzol Reagent (nitrogen) and purified using SV Total RNA Isolation System (Promega)	The hybridized microarray slides were scanned using an Agilent scanner. The relative hybridization intensities and background hybridization values were calculated using Feature Extraction Software version 9.5.1.1 (Agilent Technologies).
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	nucleic acid extraction protocol	array scanning protocol
Experimental Factor Name	treatment	organism part				
Experimental Factor Type	treatment	organism part				
Comment[SecondaryAccession]	GSE70146					
Comment[GEOReleaseDate]	2015-12-31					
Comment[ArrayExpressSubmissionDate]	2015-06-22					
Comment[GEOLastUpdateDate]	2016-01-01					
Comment[AEExperimentType]	transcription profiling by array					
SDRF File	E-GEOD-70146.sdrf.txt