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| Status |
Public on Jun 17, 2009 |
| Title |
Gene expression in macrophages stimulated with LPS in conditions allowing or inhibiting NFAT activation |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Macrophages and dendritic cells (DCs) differently contribute to the generation of coordinated immune system responses against infectious agents. 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. By contrast, following antigen recognition, macrophages initiate first the inflammatory process and then switch to an anti-inflammatory phenotype for the restoration of tissue homeostasis. Following lipopolysaccharide (LPS)-stimulation the initiation of the apoptotic pathway in DCs is due the activation of NFAT proteins. 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. The initiation of this pathway is independent of TLR4 engagement, and dependent exclusively on CD14. We asked whether macrophage survival after LPS encounter was due to their inability to activate the Ca2+ pathway.
As matter of fact, bone marrow-derived macrophages were unable to mobilize Ca2+ and to translocate NFAT to the. To further investigate whether the Ca2+-NFAT pathway played any role in LPS-stimulated macrophages, we performed a comparative kinetic microarray analysis in conditions allowing or inhibiting NFAT activation. We show here that no modulation of gene expression can be attributed to NFAT.
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| Overall design |
Gene expression analyses were performed using Affymetrix GeneChips in the following groups of murine macrophaghes: 1) CD14-deficient macrophages stimulated with LPS; 2) wt macrophages stimulated with LPS in presence of EGTA; 3) wt macrophages stimulated with LPS. The following kinetic time points were examined: 0, 6 and 24 hours following LPS activation. 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.
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| Contributor(s) |
Granucci F, Zanoni I, Ostuni R |
| Citation(s) |
19525933 |
| Submission date |
Apr 17, 2009 |
| Last update date |
May 04, 2018 |
| Contact name |
Francesca Granucci |
| E-mail(s) |
francesca.granucci@unimib.it
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| Phone |
+390264483553
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| Fax |
+390264483552
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| Organization name |
University of Milano-Bicocca
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| Street address |
Piazza della Scienza 2
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| City |
Milan |
| ZIP/Postal code |
20126 |
| Country |
Italy |
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| Platforms (1) |
| GPL8321 |
[Mouse430A_2] Affymetrix Mouse Genome 430A 2.0 Array |
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| Samples (8)
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| GSM393509 |
wild type macrophages unstimulated |
| GSM393510 |
wild type macrophages stimulated with LPS for 6 hours |
| GSM393512 |
wild type macrophages stimulated with LPS for 24 hours |
| GSM393513 |
wild type macrophages stimulated with LPS for 6 hours in presence of EGTA |
| GSM393515 |
wild type macrophages stimulated with LPS for 24 hours in presence of EGTA |
| GSM393516 |
CD14-deficient macrophages unstimulated |
| GSM393518 |
CD14-deficient macrophages stimulated with LPS for 6 hours |
| GSM393519 |
CD14-deficient macrophages stimulated with LPS for 24 hours |
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| This SubSeries is part of SuperSeries: |
| GSE15759 |
NFAT-mediated gene expression response to LPS in murine dendritic cells and macrophages |
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| Relations |
| BioProject |
PRJNA122995 |
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