Alternatively activated macrophages derived from monocytes and tissue macrophages are phenotypically and functionally distinct
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ABSTRACT: Macrophages adopt an alternatively activated phenotype (AAM) when activated by IL-4. While these AAMs can be derived from local proliferation of resident tissue macrophages or recruited inflammatory monocytes, it is not known whether these different AAMs are phenotypically and functionally distinct. Using microarray analysis of gene expression, we demonstrated that while both monocyte- and tissue-derived AAM expressed high levels of Arg1, Chi3l3 and Retnla, only monocyte-derived AAM upregulated Raldh2 and PD-L2. The distinction between monocyte- and tissue-derived macrophages can, therefore, be made using different phenotypic markers. Gene expression profiling analysis with whole genome microarray of tissue-derived and monocyte-derived macrophages, induced using IL-4c and/or 4% thioglycollate.
Project description:Macrophages adopt an alternatively activated phenotype (AAM) when activated by IL-4. While these AAMs can be derived from local proliferation of resident tissue macrophages or recruited inflammatory monocytes, it is not known whether these different AAMs are phenotypically and functionally distinct. Using microarray analysis of gene expression, we demonstrated that while both monocyte- and tissue-derived AAM expressed high levels of Arg1, Chi3l3 and Retnla, only monocyte-derived AAM upregulated Raldh2 and PD-L2. The distinction between monocyte- and tissue-derived macrophages can, therefore, be made using different phenotypic markers.
Project description:We used microarrays to find Stat6 dependent genes in control and IL-4 exposed bone marrow derived macrophages. Alternatively activated macrophages (AAM) accumulate in tissues during Th2-associated immune responses like helminth infections and allergic disorders. These cells possess potent inhibitory activity against T cells. The differentiation of AAM depends on IL-4/IL-13-mediated activation of the transcription factor Stat6. Stat6 is also required in AAM to induce several genes, such as YM1, FIZZ1 and Arginase1. Macrophages were differentiated from bone marrow cells in supplemented RPMI 1640 for 7 days in the presence of 10% supernatant from the M-CSF producing fibroblast cell line L929. After removal of nonadherent cells, macrophages were detached using 5mM EDTA treatment, washed, counted, and replated at a density of 10x6 cells/mL. Macrophages were cultured for 24 hours in medium (control) or in the presence of IL-4 (10mg/mL). F4/80 positive macrophages were sorted with a purity of 99% and RNA was isolated (Trizol/Chloroform) according to the manufacturer's instructions. RNA was labeled (biotin), fragmented, and hybridized to the arrays according to the manufacturer's recommendations.
Project description:We used microarrays to find Stat6 dependent genes in control and IL-4 exposed bone marrow derived macrophages. Alternatively activated macrophages (AAM) accumulate in tissues during Th2-associated immune responses like helminth infections and allergic disorders. These cells possess potent inhibitory activity against T cells. The differentiation of AAM depends on IL-4/IL-13-mediated activation of the transcription factor Stat6. Stat6 is also required in AAM to induce several genes, such as YM1, FIZZ1 and Arginase1.
Project description:Analysis of alternative activation of macrophages at gene expression level. The study forms part of a wider study where we compare the effects of IL-4 in different human and mouse macrophages. Our results support the notion that in vitro culture conditions greatly affect the macrophage response to IL-4. Total RNA obtained from Thioglycollate and Biogel elicited peritoneal macrophages exposed to 20 ng/ml of IL-4 for 18 hours. Biogel and thioglycollate elicited macrophages were stimulated with the Th2 cytokine IL-4, for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Similar to asthma, nematode infections are commonly associated with production of Th2 cytokines and hyporesponsive T cells. This T cell phenotype has been reported to be induced by IL-4 dependent macrophages termed alternatively activated macrophages (AAM). Additionally, AAM have been implicated in several pulmonary diseases, including allergic asthma. This study examines the potential importance of AAM as immune regulatory cells in both infectious and noninfectious disease contexts. http://www.hopkins-genomics.org/asthma/asthma006/index.html Keywords: other
Project description:This SuperSeries is composed of the following subset Series: GSE19482: Transcriptional responses of human monocyte-derived macrophages (HMDM) to lipopolysaccharide (LPS) GSE19490: Transcriptional responses of mouse BMM and TEPM to lipopolysaccharide (LPS) GSE19765: Transcriptional responses of human monocyte-derived macrophages (HMDM) to lipopolysaccharide (LPS) - Illumina arrays GSE19766: Transcriptional responses of mouse bone marrow-derived macrophages (BMM) to lipopolysaccharide (LPS) - Illumina arrays Refer to individual Series
Project description:Similar to asthma, nematode infections are commonly associated with production of Th2 cytokines and hyporesponsive T cells. This T cell phenotype has been reported to be induced by IL-4 dependent macrophages termed alternatively activated macrophages (AAM). Additionally, AAM have been implicated in several pulmonary diseases, including allergic asthma. This study examines the potential importance of AAM as immune regulatory cells in both infectious and noninfectious disease contexts. http://www.hopkins-genomics.org/asthma/asthma006/index.html
Project description:microRNAs are small non-coding RNAs that regulate gene expression at a post-translational level, and play a crucial role in the development of cells of the immune system. Macrophages are essential for generating inflammatory reactions upon tissue damage and encountering of invading pathogens, yet modulation of their immune responses is critical for maintaining tissue homeostasis. Macrophages can present different phenotypes, depending on the cytokine environment they encounter in the affected tissues. In this study, we have identified expression signatures of miRNAs that are differentially regulated during maturation of monocytes and polarization of macrophages by cytokines. We present a comprehensive characterization of miRNA expression in human monocytes and M1, M2a and M2c polarized macrophages, using next-generation Sequencing by Oligonucleotide Ligation and Detection (SOLiD). We have analyzed freshly isolated human monocytes and 5 day monocyte-derived macrophages unstimulated, or stimulated with IFNgamma+TNFalpha (M1), IL-4 (M2a) or IL-10 (M2c).
Project description:The PPARD ligand response (agonist and inverse agonist) in monocyte derived macrophages from 3 healthy donors was assesed via RNAseq.