Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression analysis of IL-4 primed bone marrow macrophages isolated from Irf4-deficient or heterozygous mice

ABSTRACT: To analyse the Irf4-dependent transcriptional changes of mouse bone marrow-derived macrophages (BMM) in response to IL-4, we have employed whole genome microarray expression profiling. For this purpose, bone marrow cells were isolated from 8 to 12 weeks old Irf4-deficient or heterozygous mice and cultured in the presence of the macrophage colony-stimulating factor (M-CSF) . After seven days of culture, IL-4 was added for 4 and 18 hours. Keywords: Mouse strain comparision; Gene expression profiling IL-4 induced gene expression was investigated in mouse bone marrow-derived macrophages (BMM) of Irf4-deficient or heterozygous mice. BMM were incubated with mouse recombinant IL-4 for 4 or 18 hours or without for 18 hours. Three independent experiments were performed at each time point (mock, 4 and 18 hours) using littermates for each experiment.

ORGANISM(S): Mus musculus

SUBMITTER: Michael Rehli

PROVIDER: E-GEOD-21853 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Interferon (IFN)γ and interleukin (IL)-4 are central regulators of T helper 1 (Th1) and T helper 2 (Th2) immune responses, respectively. Both cytokines have a major impact on macrophage phenotypes: IFNγ–priming and subsequent TLR4 activation induces so called classically activated macrophages that are characterized by pronounced pro-inflammatory responses, whereas IL-4–treated macrophages, commonly called alternatively activated, are known to develop enhanced capacity for endocytosis, antigen presentation, and tissue repair and are generally considered anti-inflammatory. Considering IL-4 as priming rather than activating stimulus, we now compared the TLR4–dependent global gene activation program in IFNγ– versus IL-4–pretreated mouse macrophages, which has rarely been studied so far. Although both cytokines frequently induced opposing effects on gene transcription, the subsequent activation of bone marrow-derived macrophages by lipopolysaccharide (LPS) produced a strong, priming dependent pro-inflammatory response in both macrophage types. For example, the production of key pro-inflammatory cytokines IL-6 and IL-12 was significantly higher in IL-4– versus IFNγ–primed macrophages and several cytokine genes, including Il19, Ccl17, Ccl22, Ccl24 and Cxcl5, were preferentially induced in alternatively primed and LPS activated mouse macrophages. In a subset of genes, including IL12a, IFNγ priming was actually found to suppress LPS–induced gene expression in a Stat1–dependent manner. Our data suggest that IL-4–priming is not per se anti-inflammatory but generates a macrophage that is “tissue protective” but still capable of mounting a strong inflammatory response after TLR4–dependent activation. Keywords: Gene expression profiling Gene expression was investigated in mouse bone marrow-derived macrophages (BMM). On day 7, BMM were stimulated with either IL-4 or IFNγ overnight (18h in total). LPS treatment was performed in primed and unprimed macrophages 4 h prior to harvesting. At least three independent experiments were performed for each condition.

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Gene expression analysis of IL-4 primed bone marrow macrophages isolated from Irf4-deficient or heterozygous mice

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