Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse bone marrow-derived dendritic cells stimilated with LPS in the presence or absence of cAMP

ABSTRACT: genes regualted by LPS or LPS+cAMP stimulation in BMDCs; We used microarrays to identify genes that up-regulated by LPS+cAMP compared with just LPS. Experiment Overall Design: BMDCs were stimilated with LPS (10 ng/ml) in the presence or absence of cAMP (100 microM) for 3h. Specifically up-regualted gene by cMP was identified.

ORGANISM(S): Mus musculus

SUBMITTER: Akihiko Yoshimura

PROVIDER: E-GEOD-16761 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Dendritic cells (DCs) are a special class of leukocytes able to activate both innate and adaptive immune responses. They interact with microbes through germline-encoded pattern-recognition receptors (PRRs), which recognize molecular patterns expressed by various microorganisms. Upon antigen binding, PRRs instruct DCs for the appropriate priming of natural killer cells, followed by specific T-cell responses. Once completed the effector phase, DCs reach the terminal differentiation stage and eventually die by apoptosis. We have observed that following lipopolysaccharide (LPS)-stimulation the initiation of the apoptotic pathway in DCs is due the activation of NFAT proteins. Indeed, LPS induces Src-family kinase and phospholipase C (PLC)Î³2 activation, influx of extracellular Ca2+ and calcineurin-dependent nuclear NFAT translocation. The initiation of this pathway is independent of TLR4 engagement, and dependent exclusively on CD14. According with this observation CD14-deficient DCs do not die following LPS stimulation. Nevertheless, CD14-deficient DC death following LPS activation can be restored by co-stimulating DCs with LPS and thapsigargin. Thapsigargin empties the intracellular calcium stores by blocking calcium pumping into the sarcoplasmic and endoplasmic reticulum and thereby activates plasma membrane calcium channels. This, in turn, allows an influx of calcium into the cytosol and NFAT activation. To identify the NFAT controlled apoptosis genes in LPS activated DCs we have performed a kinetic microarray analysis (0, 48 and 60 h) in conditions allowing or inhibiting NFAT activation. Four genes have been selected: Nur77, Gadd45g, Ddit3/Gadd153/Chop-10 and Tia1. To identify apoptosis genes selectively modulated by NFAT, a comparative kinetic (time points 0, 48 and 60 h) microarray analysis was performed in the following conditions: 1) wild type bone marrow derived DCs (wtBMDCs) stimulated with LPS; 2) CD14-deficient BMDCs stimulated with LPS; 3) wtBMDCs stimulated with LPS in presence of thapsigargin; 4) CD14-deficient BMDCs stimulated with LPS in presence of thapsigargin.

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Transcription profiling of mouse bone marrow-derived dendritic cells stimilated with LPS in the presence or absence of cAMP

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