Unknown,Transcriptomics,Genomics,Proteomics

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Bone marrow macrophage (BMMF) response to IFNgR+FcgRI coincidence detection

ABSTRACT: Different cell types that make an organism use a small number of identical signaling modules to integrate external cues and elicit diverse functions. How the specificity of functional outcomes is achieved is unclear. It must lie within the context in which the external cue is perceived and integrated by the target cell. Within the innate immune system, we discovered communication between two classes of heterologous receptors designed to integrate external cues into context-specific response. Using IFNgR as an example, we describe the structural and functional collaboration between cytokine receptors and the ITAM signaling module, in this case the Fcg-chain at the plasma membrane of phagocytes. Receptor coupling allowed IFNg to modify FcgRI responses during IgG-mediated phagocytosis and to specify cell autonomous antimicrobial functions. BMMF were left unstimulated or were stimulated in duplicates for 6 hrs with IFNÎ³, IgG2a-opsonised SRBC or both. cDNA was analyzed using 8-sample Mouse Ref-8 v2.0 Illumina array. Expressed genes were defined as those with p<0.05, and their relative expression values were calculated using unstimulated cells as calibrator. IgG2a against SRBC was produced by hybridoma TIB111 from ATCC

ORGANISM(S): Mus musculus

SUBMITTER: Ruslan Medzhitov

PROVIDER: E-GEOD-54811 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Macrophages rapidly engulf apoptotic cells to limit the release of noxious cellular contents and to restrict autoimmune responses against self antigens. Although factors participating in recognition and engulfment of apoptotic cells have been identified, the transcriptional basis for the sensing and silently disposing of apoptotic cells is unknown. Here we show that peroxisome proliferator activated receptor delta (PPARdelta) is induced when macrophages engulf apoptotic cells and functions as a transcriptional sensor of dying cells. Genetic deletion of PPARdelta decreases expression of opsonins, such as C1qb, resulting in impairment of apoptotic cell clearance and reduction in anti-inflammatory cytokine production. This increases autoantibody production and predisposes global and macrophage-specific PPARdelta deficient mice to autoimmune kidney disease, a phenotype resembling the human disease systemic lupus erythematosus. Thus, PPARdelta plays a pivotal role in orchestrating the timely disposal of apoptotic cells by macrophages, ensuring that tolerance to self is maintained. HoxA9 immortalized ES cell derived myeloid precursors were created and differentiated as in Odegaard et al, 2007. After expansion, macrophages were differentiated for 8 days in the presence of MCSF in 20% L929 conditioned media with 2.5% bovine serum albumin in low glucose media. Macrophages were harvested for RNA using Qiagen's RNeasy Kit. A common reference was generated using e17.5 C57Bl/6 embryos. 30ug of either reference or macrophage RNA was labeled with Cy3 and Cy5 (Amersham) and hybridized to Stanford Functional Genomics Facility mouse cDNA microarrays for 16 hours. The arrays were washed and scanned on an Agilent G2505A ChipScan v. A.6.3.1. Florescence values were extracted using GenePix Pro v. 5.0.0.51. The data were normalized using regression correlation. Subsequently, missing values were imputed and significantly differentially expressed genes were found with Significance Analysis of Microarrays. For the Nature Medicine manuscript, significantly differentially expressed genes including genes involved in the process of uptake of apoptotic cells were extracted and clustered by complete linkage using Cluster and visualized using TreeView. Key to genotypes: "J1 untreated" are WT differentiated macrophages; "18 Untreated" are PPARd-/- differentiated macrophages Genotype: Wild type (WT) or PPAR delta minus (PPARd -/-) macrophages genetic_modification_design

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Bone marrow macrophage (BMMF) response to IFNgR+FcgRI coincidence detection

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