The Cytokines Interleukin-1 and Interferon-gamma Differentially Program Beta-glucanM-bM-^@M-^Sactivated Dendritic Cells via IkappaB-zeta Modulation
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Marco Cardone
PROVIDER: E-GEOD-42189 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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2008-06-13 | E-GEOD-5151 | biostudies-arrayexpress
Project description:IL-17A is a pro-inflammatory cytokine that promotes host defense against infections and contributes to the pathogenesis of chronic inflammatory diseases. Dendritic cells (DC) are antigen-presenting cells responsible for adaptive immune responses. Here, we report that IL-17A induces intense remodeling of lipid metabolism in human monocyte-derived DC, as revealed by microarrays analysis. In particular NR1H3/LXR-a and its target genes were significantly upregulated in response to IL-17A. IL-17A induced accumulation of Oil Red O-positive lipid droplets in DC leading to the generation of lipid-laden DC. A lipidomic study established that all the analyzed lipid species, i.e phospholipids, cholesterol, triglycerides, cholesteryl esters were elevated in IL-17A-treated DC. The increased expression of membrane lipid transporters in IL-17A-treated DC as well as their enhanced ability to uptake the fatty acid Bodipy-FL-C16 suggested that lipid uptake was the main mechanism responsible for lipid accumulation in response to IL-17A. IL-17A-induced lipid laden DC were able to stimulate allogeneic T cell proliferation in vitro as efficiently as untreated DC, indicating that IL-17A-treated DC are potently immunogenic. This study, encompassed in the field of immunometabolism, points out for the first time IL-17A as a modulator of lipid metabolism in DC and provides a rationale to delineate the importance of lipid-laden DC in IL-17A-related inflammatory diseases. We used microarrays analysis to understand the impact of IL-17A on human monocyte-derived human dendritic cells. We found overexpression of many genes involved in lipid metabolism in IL-17A-treated dendritic cells compared to untreated dendritic cells. In particular NR1H3/LXR-a and its target genes were significantly upregulated in response to IL-17A. IL-17A induced accumulation of Oil Red O-positive lipid droplets in DC leading to the generation of lipid-laden DC. A lipidomic study established that all the analyzed lipid species, i.e phospholipids, cholesterol, triglycerides, cholesteryl esters were elevated in IL-17A-treated DC. The increased expression of membrane lipid transporters in IL-17A-treated DC as well as their enhanced ability to uptake the fatty acid Bodipy-FL-C16 suggested that lipid uptake was the main mechanism responsible for lipid accumulation in response to IL-17A. IL-17A-induced lipid laden DC were able to stimulate allogeneic T cell proliferation in vitro as efficiently as untreated DC, indicating that IL-17A-treated DC are potently immunogenic. This study, encompassed in the field of immunometabolism, points out for the first time IL-17A as a modulator of lipid metabolism in DC and provides a rationale to delineate the importance of lipid-laden DC in IL-17A-related inflammatory diseases. RNA was extracted from untreated in vitro-generated DC at day 0 (DC, 4 biological replicates ) or DC cultured for 12 days with IL-17A, in the absence or presence of IFN-g (DC-17 and DC-G17, 5 biological replicates)
2015-03-31 | E-GEOD-53163 | biostudies-arrayexpress
Project description:Generation of human monocyte-derived DCs Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density centrifugation from buffy coats or whole blood of healthy donors and resuspended in serum-free medium (CellGro, CellGenix GmbH, Freiburg, Germany), supplemented with penicillin (50U/ml)/streptomycin (50?g/ml). Cells were plated at a density of 1-1,25 e6/cm2 in T-25 flasks and allowed to adhere for 2h at 37oC. Non-adherent cells were removed by washing twice with PBS and frozen as T cell source for stimulation experiments. Adherent monocytes were differentiated into immature DC with 1000U/ml GM-CSF and 50ng/ml IL-4 (""im DC""). Unless stated otherwise, cytokines were purchased from CellGenix GmbH, Freiburg, Germany. Cytokines were replenished on day 3. Maturation was achieved by incubation with either IL-1?; (10ng/ml), IL-6 (1000U/ml), TNF?; (10ng/ml) and PGE2 (1?g/ml, ICN, Aurora, Ohio, USA) (""PGE2-DC"") or IL-1?; (25ng/ml), IFN?; (3000U/ml, Schering-Plough, Brussels, Belgium), IFN?; (1000U/ml Strathmann Biotech AG), TNF?; (50ng/ml) and poly(I:C) (a synthetic, double-stranded RNA, 20ng/ml, Sigma-Aldrich, Taufkirchen, Germany) (""poly(I:C)-DC"") on day 6 for 48 hours Total RNA was isolated from DC before and after cytokine maturation
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