Transcriptional profiling in macrophages in stimulated and unstimulated conditions.
Ontology highlight
ABSTRACT: Transcriptomes from macrophages at three stages were examined: a) Non-stimulated, b)Stimulated by Interleukin 4, c)Stimulated by LPS and Interferon gamma. Four biological replicate of each experiment were performed.
Project description:Extracellular membrane vesicles (MVs) are powerful biomarkers in several pathological processes. The potential advantage of MVs relays on the assumption that their content reflects processes ongoing in pathologically relevant cell types. Using microarrays, we performed transcriptional profiling in stimulated (M1 and M2) and unstimulated conditions.
Project description:Understanding the respective role of microglia and infiltrating monocytes in neuroinflammatory conditions has recently seemed possible by the identification of a specific microglia signature. Here instead we provide evidence that peripheral macrophages may express some of the most commonly described microglia markers at some developmental stages or pathological conditions, in particular during chronic neuroinflammation. Further, our data support the hypothesis about phenotypic plasticity and convergence among distinct myeloid cells so that they may act as a functional unit rather than as different entities, boosting their mutual functions in different phases of disease. This holds relevant implications in the view of the growing use of myeloid cell therapies to treat brain disease in humans.
Project description:This SuperSeries is composed of the following subset Series: GSE31529: Genome-wide binding of STAT3 in peritoneal macrophages GSE31530: Transcriptome changes in IL-10 treated peritoneal macrophages Refer to individual Series
Project description:Mast cells are hematopoietic cells that reside preferentially in tissues exposed to internal and external environments. Mast cells sense immunological, inflammatory and environmental stimuli, and can be activated to release granules and generate inflammatory mediators. Mast cell-derived products confer protection against snake venoms and some parasite infections. Aberrant activation of mast cells is a major contributor to human pathology, including allergy, asthma and adverse drug reactions. Their strict tissue location has largely impeded the isolation of large numbers of primary mast cell for further analysis. To better understand the biology of mast cells, we analyzed the proteome of primary mouse mast cells by quantitative mass spectrometry
Project description:To try to identify the mechanism of STAT3âs indirect action we have used a genomic approach to map the binding sites of STAT3 within the genome and also used RNA-seq technology to map the changes in RNA expression and transcript isoforms in response to IL-10. Examination of STAT3 binding by ChIP-seq in Unstimulated and IL-10 treated peritoneal exudate macrophages purified from mice. We sequenced anti-STAT3 ChIP-seq as well as corresponding control (Input) libraries
Project description:Toll-like receptor and RIG-I-like receptor classs may evoke or instruct a distinct type of response that is more reflective of the pathogen encountered. Although this issue may be critical to a better understanding of the regulation of immune responses to microbial infections, it has not been addressed through a direct, side-by-side comparison of the two receptor classes. To address this, we performed microarray analysis of mRNAs from peritoneal macrophages stimulated with a synthetic B-form DNA, poly(dA-dT)M-cM-^CM-;poly(dT-dA) ( B-DNA) or a CpG-B oligonucleotide, which respectively activates RLRs or TLR9, and identified B-DNA inducible genes and CpG-B inducible genes. Peritoneal macrophages from B6 mice were stimulated with B-DNA (10M-NM-<g/ml) or CpG-B(3M-NM-<M) for 4 hous, and then subjected to microarray analysis.
Project description:F4/80+ macrophages treated with TGFb2 are potently tolerogenic. Our understanding of the molecular mechanisms mediating the development of these tolerogenic properties is incomplete. We used microarray analysis to identify molecules that are involved in the tolerogenic mechanisms in murine TGFb-treated macrophages. Thioglycolate-elicited peritoneal exudate cells (F4/80+ macrophages) were treated with either PBS or TGFb2 overnight then total RNA extracted and subjected to microarray analysis.
Project description:Alternatively activated macrophages (AAMφs) play important roles in a number of Th2 driven pathologies including asthma and allergy, and a number of parasitic infections. Our studies, and those of others, investigating the pathologies associated with infection with the helminth Schistosoma japonicum implicate a role for AAMφs in fibrosis and immunomodulation.. In the present study we show that S. japonicum-secreted egg antigens are able to induce the alternative activation of macrophages as characterised by the induction of Chi3l3 and Arg1 expression. Retnla, another common marker of AAMφs, was not consistently induced in these macrophages suggesting that the specific function of these cells may differ to those induced by S. mansoni and other parasites. Closer examination of the gene expression profile and functionality of these cells identified pathways independent of Retnla expression that could be important for their immunomodulatory activity such as modulating expression of T-cell co-stimulatory molecules and chemokines. In vivo generated S. japonicum soluble egg antigen stimulated AAMφs also exhibited a reduction in their phagocytic ability likely related to the induction of IL4 and decreased expression of cell surface receptors. Additionally these macrophages exhibited reduced expression of Toll-like receptors (TLRs) and an associated reduction in responsiveness to stimulation with TLR ligands. We did not observe pathways that would suggest that AAMφs have a direct profibrotic activity. Taken together, these data describe a mechanism by which alternative activation of macrophages may be induced during S. japonicum infection and highlight the importance of the context of activation in directing AAMφ phenotype and function. The gene expression profile of Schistosoma japonicum soluble egg antigen (SEA)-stimulated macrophages was compared with that of PBS stimulated controls. Macrophages were isolated from the peritoneal cavity of BALB/c mice (n=6 per group) stimulated by intraperitoneal injection with SEA or PBS. The macrophages were pooled and RNA was extracted from these cells. Microarray analysis was performed on cRNA synthesised from total RNA derived from these macrophages. The experiment was performed twice creating two biological replicates. Fold-change (relative to the respective PBS control) reported in supplementary file linked below.
Project description:This study dermined the temporal gene expression profile of myeloid subpopulations recruited to the peritoneal cavity to encapsulate implanted foreign material. Sterile foreign objects were inserted into the peritoneal cavities of MacGreen mice (in which the Csf1r promoter directs myeloid-specific EGFP expression). At various time-points post-surgery (days 2, 4, 7, 14), mice were euthanased, and peritoneal exudate cells removed by lavage. Peritoneal exudate cells from MacGreen mice that had not received implants were used as controls (day 0; 'unstimulated'). Free-floating objects encapsulated with tissue were removed from the peritoneal cavities of different mice at days 7, 14, 21, 28 days, and single cell suspensions obtained by collagenase digestion. Single-cell suspensions from peritoneal exudate or tissue capsules were separated on the basis of size/granularity and EGFP fluorescence using FacsVantage SE Diva. Total RNA was extracted from FACS-sorted EGFP-hi cells.
Project description:microRNA transcriptome data from wild type and Gata6-deficient tissue resident peritoneal macrophages. Tissue resident macrophages are notoriously heterogeneous, exhibiting discrete phenotypes as a consequence of tissue- and micro-anatomical niche-specific functions, but the molecular basis for this is not understood. Gata6 itself has been shown to be a target of multiple miR. However, microRNA transcriptome and its dependence on tissue-specific macrophage programming, such as effected by GATA6, has not been explored. We used microRNA sequencing to determine the patterns of microRNA expression in peritoneal resident macrophages at homeostasis in the absence of GATA-6 against wild type.