Mus musculus
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ABSTRACT:
PROVIDER: PRJNA103841 | ENA |
REPOSITORIES: ENA
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Project description:The receptors engaged during phagocytic particle uptake determine the signaling events that occur during phagosome formation and maturation. However, pathogens generally have multiple ligands, making it difficult to dissect the roles of individual receptors in these processes. Here, we used latex beads coupled to single ligands, focusing on IgG, mannan, LPS and avidin, and monitored phagocytic uptake rates, phago-lysosomal fusion events, macrophage gene expression and the proteomic composition of isolated phagosomes. The pattern of gene expression and the protein composition of isolated phagosomes showed that each bead ligand altered a distinct pattern of genes and led to a different composition of phagosomes. These data argue that activation of each receptor initiates a specific signature of signaling events that last many hours and influences several phagocytosis functions. All samples and controls were carried out in triplicates. J774.A1 cells (mouse macrophage-like cell line) were seeded onto 6-well plates one day before the experiment. Subsequently, cells were incubated with serum-free DME medium containing 0.01 % latex beads of 1 µm diameter coupled to the following ligands: avidin (Av), the Fc fragment of mouse IgG (Fc), lipopolysaccharides from Klebsiella pneumoniae (LPS) and mannan from Saccharomyces cerevisae (Man) for 30 and 60 minutes. After incubation, samples as well as untreated controls were washed twice in PBS and total RNA was extracted using RNeasy kit (Qiagen) following manufacturer's instructions. Each sample was hybridized to CodeLink Mouse whole genome bioarray slides (Amersham). This study was intended to analyze the role of receptor-ligand interactions on phagosome maturation and gene expression after receptor-mediated phagocytosis in macrophages. CodeLink EXP v4.0-processed data are represented in the Sample tables, GeneSpring-processed data are linked as a supplementary files to the matrix table, additional results available as a supplementary file on the Series record. http://www.biologie.uni-rostock.de/tierphysiologie/agkuznetsov.html
2010-05-05 | E-GEOD-17761 | biostudies-arrayexpress
Project description:The type of phagocytic receptor modulates gene expression and phagosome functions in macrophages
| PRJNA118305 | ENA
Project description:Tumor Progression Locus 2 (TPL-2) kinase mediates Toll-like Receptor (TLR) activation of ERK1/2 and p38 MAP kinases in myeloid cells to modulate expression of key cytokines in innate immunity. This study identified a novel MAP kinase-independent regulatory function for TPL-2 in phagosome maturation, an essential process for killing of phagocytosed microbes. TPL-2 catalytic activity was demonstrated to induce phagosome acidification and proteolysis in primary mouse and human macrophages following uptake of latex beads. Quantitative proteomics revealed that blocking TPL-2 catalytic activity significantly altered the protein composition of phagosomes, particularly reducing the abundance of V-ATPase proton pump subunits. Furthermore, TPL-2 stimulated the phosphorylation of DMXL1, a critical regulator of V-ATPases, to induce phagosome acidification. Consistent with these results, TPL-2 catalytic activity was required for phagosome maturation and the efficient killing of Staphylococcus aureus following phagocytic uptake by macrophages. These results indicate that TPL-2 controls the innate immune response of macrophages to bacteria via V-ATPase induction of phagosome maturation.
2021-03-24 | PXD020247 | Pride
Project description:Tumor Progression Locus 2 (TPL-2) kinase mediates Toll-like Receptor (TLR) activation of ERK1/2 and p38-alpha MAP kinases in myeloid cells to modulate expression of key cytokines in innate immunity. This study identified a novel MAP kinase-independent regulatory function for TPL-2 in phagosome maturation, an essential process for killing of phagocytosed bacteria. TPL-2 catalytic activity was demonstrated to induce phagosome acidification and proteolysis in primary mouse and human macrophages following uptake of latex beads. Mass spectrometry analysis revealed that blocking TPL-2 catalytic activity significantly altered the protein composition of phagosomes, particularly reducing the abundance of V-ATPase proton pump subunits. Furthermore, TPL-2 was shown to stimulate the phosphorylation of DMXL1, a critical regulator of V-ATPases, to induce phagosome acidification. Consistent with these results, TPL-2 catalytic activity was required for phagosome acidification, activation of phagosome acid-sensitive cathepsins and the efficient killing of Staphylococcus aureus following phagocytic uptake by macrophages. These results indicate that TPL-2 controls the innate immune response of macrophages to bacteria via MAP kinase regulation of gene expression and V-ATPase induction of phagosome maturation.
2021-02-11 | PXD020401 | Pride
Project description:The receptors engaged during phagocytic particle uptake determine the signaling events that occur during phagosome formation and maturation. However, pathogens generally have multiple ligands, making it difficult to dissect the roles of individual receptors in these processes. Here, we used latex beads coupled to single ligands, focusing on IgG, mannan, LPS and avidin, and monitored phagocytic uptake rates, phago-lysosomal fusion events, macrophage gene expression and the proteomic composition of isolated phagosomes. The pattern of gene expression and the protein composition of isolated phagosomes showed that each bead ligand altered a distinct pattern of genes and led to a different composition of phagosomes. These data argue that activation of each receptor initiates a specific signature of signaling events that last many hours and influences several phagocytosis functions.
2009-09-10 | GSE17761 | GEO
Project description:Resident tissue macrophages (RTMs) are organ-specialized phagocytes responsible for the maintenance and protection of residing tissue. It is well established that tissue diversity is reflected by the heterogeneity of RTMs origin and phenotype. However, much less is known about tissue-specific phagocytic macrophage functions. Here, using quantitative proteomics approach, we identify cathepsins as key determinants of phagosome maturation in primary peritoneal, lung and brain resident macrophages. The data further uncover cathepsin K (CtsK) as a molecular marker for lung phagosomes required for intracellular protein and collagen degradation. Pharmacological blockade of CtsK activity diminished phagosomal proteolysis and collagenolysis in lung resident macrophages. Furthermore, pro-fibrotic TGF-β negatively regulated CtsK-mediated phagosomal collagen degradation independently from classical endocytic proteolytic pathways. In humans, phagosomal CtsK activity was reduced in lung COPD macrophages and lung macrophages exposed to cigarette smoke extract. Taken together, this study provides a comprehensive map of how peritoneal, lung and brain tissue environment shapes phagosomal composition of resident macrophages, revealing CtsK as a key molecular determinant of lung phagosomes contributing to phagocytic collagen clearance in lungs.
2023-01-18 | PXD033010 | Pride
Project description:Macrophages mediate the elimination of pathogens by phagocytosis resulting in the activation of specific signaling pathways that lead to the production of cytokines, chemokines and other factors. Borrelia burgdorferi, the causative agent of Lyme disease, causes a wide variety of pro-inflammatory symptoms. The proinflammatory capacity of macrophages is intimately related to the internalization of the spirochete. However, most receptors mediating this process are largely unknown. We have applyedapplied a multiomic approach, including the proteomic analysis of B. burgdorferi-containing phagosome-enriched fractions, to identify surface receptors that are involved in the phagocytic capacity of macrophages as well as their inflammatory output. Sucrose gradient protein fractions of human monocyte-derived macrophages exposed to B. burgdorferi contained the phagocytic receptor, CR3/CD14 highlighting the major role played by these proteins in spirochetal phagocytosis. Among others, Other proteins identified proteins include C-type lectins, scavenger receptors or siglecs, and contain uPAR and MARCO. We also identified the Fc gamma receptor pathway as involved both in the phagocytosis of , and TNF induction by B. burgdorferi in the absence of antibodies. The common gamma chain, FcR, mediates the phagocytosis of the spirochete, likely through Fc receptors and C-type lectins, in a process that involves Syk activation. Overall, these findings highlight the complex array of receptors involved in the phagocytic response of macrophages to B. burgdorferi.
2019-12-06 | PXD012854 | Pride
Project description:As part of a study comparing phagosome composition in wild-type Dictyostelium discoideum with mutants in two cytoskeletal/motor proteins, we compared global gene expression and proteomes between the strains to test whether differences in mRNA and protein abundance were correlated with changes in phagosomal composition. abp1 and myoK mutants in both the Ax2 and DH1 backgrounds were compared with their respective wild-type parents.Both analyses converged and confirmed the phagosome analysis (supplemental Tables). Protein differences were markedly fewer in the myoK-null mutant than in the abp1 null lysate. Proteins of known function, differential in the myoK-null lysate, were involved in remodeling of the plasma membrane and early membrane trafficking. Proteins differential in the abp1-null lysate spanned various functions at all levels of cell metabolism. Most of the proteins differential in 2D-DIGE displayed similar mutant-to-wt ratios at the mRNA level in microarrays. A prominent example of overlap is the 1.4 fold overexpression (p ??? 0.016) of Vps5, both in proteomics and microarray analyses of abp1-null cells. Like half of the proteins involved in membrane trafficking that are differential in microarray analyses of abp1-null cells, Vps5 is putatively involved in endosome-to-Golgi trafficking as a retromer sorting nexin. Therefore, one might speculate that, in addition to direct effects on phagocytic uptake, absence of Abp1 has an impact on endosome-to-Golgi trafficking.
2012-02-06 | E-MTAB-978 | biostudies-arrayexpress
Project description:The receptors engaged during phagocytic particle uptake determine the signalling events that occur during phagosome formation and maturation.However, pathogens generally have multiple ligands, making it difficult to dissect the roles of individual receptors in these processes. Here,we used latex beads coupled to single ligands, focusing primarily on IgG, mannan and avidin, and for some experiments on LPS, and monitored 1. phagocytic uptake rates, 2. fusion with lysosomal compartments, 3.macrophage gene expression 4. proteomic composition of isolated phagosomes and 5. the binding of phagosomes to actin filaments. Phagosomes formed in response to mannan fused with lysosomes earlier after uptake than the ones formed by the other ligands. The pattern of gene expression and the protein composition of isolated phagosomes showed that each bead ligand upregulated a distinct pattern of genes and led to a different composition of phagosomes. Furthermore, isolated phagosomes also exhibited differences in binding to F-actin in vitro.These data argue that activation of each receptor initiates a specific signature of signalling events that last many hours and influences several phagocytosis functions. Keywords: receptor-ligand interactions
2007-12-12 | GSE9859 | GEO
Project description:The enteric protozoan parasite Entamoeba histolytica have high phagocytic ability. Phagocytosis is also important for the pathogenicity of this parasite; molecular mechanisms of phagocytosis and phagosome maturation is focused. Atg8 is well studied autophagy marker protein. We previously shown that E. histolytica Atg8 translocate to nascent trogosomes and expression silencing of the Atg8 caused retardation of phagosome acidification. To investigate how Atg8 regulates phagosome maturation, here we conducted proteome analysis of phagosomes isolated from an E. histolytica strain in which atg8 gene expression are silenced (atg8 gene silenced, atg8-gs) and its mock control strain (transfected with psAP2-Gunma).
2022-10-20 | PXD034376 | Pride