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NRROS negatively regulates ROS in phagocytes during host defense and autoimmunity

ABSTRACT: Production of reactive oxygen species (ROS) is one of the important antimicrobial mechanisms of phagocytic cells. Enhanced oxidative burst requires these cells to be primed with agents such as IFNg and LPS with a synergistic effect of these agents on the level of the burst. However, excessive ROS generation will lead to tissue damage and has been implicated in a variety of inflammatory and autoimmune disease. Therefore, this process needs to be tightly regulated. In order to understand the genes regulating this process, we will treat bone marrow derived macrophages with above mentioned priming agents and study the gene expression. We used microarrays to determine the changes in gene expression that occur in bone marrow derived macrophages after treatment with IFNg, LPS, or a combination of IFNg and LPS

ORGANISM(S): Mus musculus

PROVIDER: GSE53986 | GEO | 2014/03/31

SECONDARY ACCESSION(S): PRJNA234284

REPOSITORIES: GEO

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NRROS negatively regulates ROS in phagocytes during host defense and autoimmunity

Project description:Production of reactive oxygen species (ROS) is one of the important antimicrobial mechanisms of phagocytic cells. Enhanced oxidative burst requires these cells to be primed with agents such as IFNg and LPS with a synergistic effect of these agents on the level of the burst. However, excessive ROS generation will lead to tissue damage and has been implicated in a variety of inflammatory and autoimmune disease. Therefore, this process needs to be tightly regulated. In order to understand the genes regulating this process, we will treat bone marrow derived macrophages with above mentioned priming agents and study the gene expression. We used microarrays to determine the changes in gene expression that occur in bone marrow derived macrophages after treatment with IFNg, LPS, or a combination of IFNg and LPS. Four condition experiment; Biological replicates: four replicates per condition

2014-03-31 | E-GEOD-53986 | biostudies-arrayexpress

Interaction between ROS and miR-451

Project description:Reactive Oxygen Species (ROS) could be a stress factor that affects microRNA regulation and function in macrophages. The production of microRNAs (miRNA) is influenced by various stimuli, including environmental stresses. We hypothesized that ROS-associated stress could regulate macrophage miRNA synthesis. p47phox-/- mice have deficient NADPH oxidase activity resulting in decreased ROS production. We cultured bone marrow-derived macrophages (BMDM) from wild type (WT) and p47phox-/- mice and profiled miRNA expression using microarrays. The microarray data reveals that there are differences in the expression levels of different miRs, and our studies suggest functional crosstalk between ROS and miR-451 in the regulation of macrophage oxidant stress. Mouse bone marrow-derived macrophages (BMDMs) were obtained from WT (wild type) and p47phox-/- mice. MicroRNAs were isolated by using the mirVana miRNA kit, and a TaqMan rodent microRNA array (consisting of Megaplex RT Primers, Rodent Pool-A, Applied Biosystems) was used for microarray. The array enables quantitation of the expression levels of up to 380 microRNAs and controls. Rodent Pool A contains reverse transcription (RT) primers for 335 and 238 unique microRNAs for mouse and rat, respectively, plus 4 species-specific controls. The data were analyzed on RQ manager software (Qiagen, SA Biosciences) and normalized to the endogenous controls, and analyzed for fold change of miRs in WT compared to p47phox-/-.

2014-07-09 | E-GEOD-55189 | biostudies-arrayexpress

Interaction between ROS and miR-451

2014-07-09 | GSE55189 | GEO

Delineation of diverse macrophage activation programs in response to intracellular parasites and cytokines

Project description:To provide a global perspective on the relationships between macrophage activation programs and to understand how certain pathogens circumvent them, we used transcriptional profiling by genome wide microarray analysis to compare the responses of mouse macrophages following exposure to the intracellular parasites Trypanosoma cruzi and Leishmania mexicana, the bacterial product lipopolysaccharide (LPS), and the cytokines IFNG, TNF, IFNB, IL-4, IL-10, and IL-17. We found that LPS induced a classical activation state that resembled macrophage stimulation by the Th1 cytokines IFNG and TNF. However, infection by the protozoan pathogen Leishmania mexicana produced so few transcriptional changes that the infected macrophages were almost indistinguishable from uninfected cells. Trypanosoma cruzi activated macrophages produced a transcriptional signature characterized by the induction of interferon-stimulated genes by 24 h post-infection. Despite this delayed IFN response by T. cruzi, the transcriptional response of macrophages infected by the kinetoplastid pathogens more closely resembled the transcriptional response of macrophages stimulated by the cytokines IL-4, IL-10, and IL-17 than macrophages stimulated by Th1 cytokines. Keywords: Bone marrow macrophage response to intracellular parasites and cytokines We analyzed a series MEEBO arrays on which were hybed RNA amplified from bone marrow-derived macrophages from C57BL/6 mice. Macrophages infected with L. mexicana or T. cruzi or stimulated by LPS, IFNG, IL-4, IL-10, TNF, IFNB, or IL-17 were compared to one another as well as to uninfected, unstimulated control macrophages. All experiments were performed over a 24 h timecourse with timepoints taken at 2 h, 6 h, 12 h, and 24 h.

2010-04-13 | E-GEOD-20087 | biostudies-arrayexpress

To determine the global gene expression response in murine LPS-induced macrophages treated with a bromodomain inhibitor.

Project description:Bone marrow-derived macrophages, Unstimulated DMSO Bone marrow-derived macrophages, Unstimulated + I-BET (GSK525762A) Bone marrow-derived macrophages, LPS 1h DMSO Bone marrow-derived macrophages, LPS 1h + I-BET (GSK525762A) Bone marrow-derived macrophages, LPS 2h DMSO Bone marrow-derived macrophages, LPS 2h + I-BET (GSK525762A) Bone marrow-derived macrophages, LPS 4h DMSO Bone marrow-derived macrophages, LPS 4h + I-BET (GSK525762A) LPS (100 ng/mL) was purchased from Sigma. Bone Marrow-derived macrophages (BMDMs) were differentiated from C57BL/6 bone marrow using 5 ng/mL each of recombinant M-CSF and IL-3 (Peprotech) for 7 days as described (Jeffrey et al, Nature Immunology, 2006). 2 x10^6 BMDMs were treated with DMSO or 1 μM of I-BET for 30 minutes before the addition of LPS (100 ng/mL) for 1, 2 or 4h. Unstimulated control samples were incubated with I-BET only for 1 hour. 500 ng of total RNA from 3 independent samples per group was used to prepare biotin-labeled RNA using Ambion Illumina TotalPrep RNA Amplification Kit (Applied Biosystems) and hybridized to Illumina MouseRef-8 v2.0 expression BeadChip kits. The chips were scanned using Illumina BeadArray Reader. 3 biological replicates and 4 timepoints

2010-11-10 | E-GEOD-21764 | biostudies-arrayexpress

Bone marrow-derived macrophage responses to classical activation: wild-type vs. MLL1-deficient (Lys2Cre MLL1 fx/fx)

Project description:mRNA from wild-type (Cre-) and MLL1-deficient (Cre+) BMDMs were analyzed via gene chip (Mouse Gene ST 2.1, Affymetrix) for relative expression changes. Isolated mRNA from Cre- and Cre+ BMDMs stimulated with classical activation signals (IFNg, LPS or IFNg+LPS) was analyzed using a gene chip panel of >40,000 RefSeq transcripts, and resulting fold expression was determined by analyzing quality-controlled expression values for validated probesets. Bone marrow derived macrophages from wild-type (Cre-) or MLL1-deficient (Cre+) mice were stimulated in vitro with IFNgamma (10 ng/ml), LPS (100 ng/ml) or the combination of IFNgamma+LPS for six hours. Cells were then processed in Trizol reagent for RNA extraction.

2016-06-02 | E-GEOD-82109 | biostudies-arrayexpress

To determine the global gene expression response in murine LPS-induced macrophages treated with a bromodomain inhibitor.

2010-11-10 | GSE21764 | GEO

Sema6D reverse signaling controls lipid metabolism for macrophage polarization linking mTOR to PPARγ

Project description:Purpose: The goal of this study is to compare downstream genes of Sema6D signaling in LPS plus IFNg stimulated macrophages. Methods: Bone marrow derived macrophage mRNA profiles of 7 weeks of wild type (WT) and Sema6D-/- mice were stimulated by LPS for 4 hrs. Results: According to this comparison, we found that 550 genes were downregulated in Sema6D-/- macrophages than WT macrophages in response to LPS. Conclusions: Our study represents 62 genes were supressed in both M1 and M2 Sema6D-/- macrophage than WT macrophages, suggesting of Sema6D reverse sigaling genes.

2018-05-20 | GSE99047 | GEO

C56BL6 WT or IFNARKO BMDM stimulated with different TLRs (part-V)

Project description:WT control or IFNAR deficient bone marrow derived macrophages were stimulated with IFNb, TLR3, IFNg, TLR4, and TLR4+IFNg ligands for 48 h.

2020-04-01 | ST001345 | MetabolomicsWorkbench

Single-cell response to LPS of wild-type and cohesin-deficient macrophages.

Project description:Transcription is an intermittent process that occurs in bursts. The frequency and the number of messenger RNA (mRNA) produced by these bursts determine expression levels and shape cell-to-cell variability. Productive encounters between enhancers and promoters control burst dynamics, which the cohesin complex can facilitate through loop extrusion. Cohesin is a ring-shaped complex involved in enhancer-promoter interactions. Without cohesin, bone-marrow-derived macrophages' transcriptional response to LPS is severely impaired (Cuartero et al., 2018). Single-cell RNA sequencing (scRNAseq) showed that the frequency of cells expressing inducible transcripts (as defined in Bhatt et al., 2012) was reduced in cohesion-deficient macrophages before and after LPS stimulation. In contrast, when considering only cells expressing LPS-inducible transcripts, cohesin-deficient macrophages showed reduced mean expression at baseline and late LPS-response (8h) but not in the early response (2h). These results support a model where cohesin is implicated in the control of burst frequencies upon LPS.

2022-04-17 | GSE190622 | GEO

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