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E-GEOD-15718 - Gene expression in dendritic cells stimulated with LPS in conditions allowing or inhibiting NFAT activation

Status
Submitted on 16 April 2009, released on 4 March 2010, last updated on 3 May 2014
Organism
Mus musculus
Samples (18)
Array (1)
Protocols (27)
Description
Interleukin-2 (IL-2) is one of the molecules produced by mouse dendritic cells (DCs) after stimulation by Toll like receptor (TLR) agonists. By analogy with the events following T-cell receptor (TCR) engagement leading to IL-2 production we have observed that DC stimulation with lipopolysaccharide (LPS) induces Src-family kinase and phospholipase C (PLC)γ2 activation, influx of extracellular Ca2+ and calcineurin-dependent nuclear NFAT translocation. We have also observed that the initiation of this pathway is independent of TLR4 engagement, and dependent exclusively on CD14. To determine the role of NFAT in LPS activated dendritic cells we have performed microarray analysis in conditions allowing or inhibiting NFAT activation. We show here that LPS-induced NFAT activation via CD14 is necessary to cause death of terminally differentiated DCs, an event that is essential for maintaining self-tolerance and preventing autoimmunity. Consequently, blocking this pathway in vivo causes prolonged DC survival and an increase in T cell priming capability. Gene expression analyses were performed using Affymetrix GeneChips in the following groups of murine bone marrow-derived dendritic cells: 1) CD14-deficient BMDCs stimulated with LPS; 2) wtBMDCs stimulated with LPS in presence of EGTA; 3) wtBMDCs stimulated with LPS. This experimental setting allowed us to select for effects due to Ca2+ fluxes and exclude the effects due to other causes, particularly the block of TRIF recruitment in CD14-deficient cells and the EGTA effects unrelated to Ca2+ chelation.
Experiment type
transcription profiling by array 
Contacts
Citation
CD14 regulates the dendritic cell life cycle after LPS exposure through NFAT activation. Zanoni I, Ostuni R, Capuano G, Collini M, Caccia M, Ronchi AE, Rocchetti M, Mingozzi F, Foti M, Chirico G, Costa B, Zaza A, Ricciardi-Castagnoli P, Granucci F.
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-15718.idf.txt
Sample and data relationshipE-GEOD-15718.sdrf.txt
Raw data (1)E-GEOD-15718.raw.1.zip
Processed data (1)E-GEOD-15718.processed.1.zip
Array designA-AFFY-36.adf.txt
R ExpressionSetE-GEOD-15718.eSet.r
Links