Dataset Information

Temporal and ligand effects on phagosome maturation

ABSTRACT: Systems biology analysis of temporally-resolved phagosome proteomes following uptake via key phagocytic receptors Macrophages operate at the forefront of innate immunity, and their discrimination of pathogenic versus “self” particles is critical for a number of responses including efficient pathogen killing, antigen presentation and cytokine induction. In order to efficiently phagocytose and destroy the particles, macrophages express an array of receptors to sense and internalise prey particles. In this manuscript, particles conjugated to seven ligands representing pathogen-associated molecular patterns, immune opsonins or apoptotic cell markers were inoculated to murine bone marrow-derived macrophages (BMDMs) and proteomes of isolated phagosomes were accurately quantified among conditions and across a timecourse. While we identified a clear functional differentiation over the three timepoints, only subtle differences were detected between certain ligand-phagosomes, suggesting that triggering of receptors through a single ligand has only mild effects on phagosome proteome and function. This data shows for the first time a systematic time-course analysis of BMDM phagosomes and how phagosome fate is regulated by the receptors triggered for phagocytosis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Matthias Trost

LAB HEAD: Matthias Trost

PROVIDER: PXD000772 | Pride | 2015-09-10

REPOSITORIES: Pride

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Publications

Quantitative proteome analysis of temporally resolved phagosomes following uptake via key phagocytic receptors.

Dill Brian D BD Gierlinski Marek M Härtlova Anetta A Arandilla Alba González AG Guo Manman M Clarke Rosemary G RG Trost Matthias M

Molecular & cellular proteomics : MCP 20150309 5

Macrophages operate at the forefront of innate immunity and their discrimination of foreign versus "self" particles is critical for a number of responses including efficient pathogen killing, antigen presentation, and cytokine induction. In order to efficiently destroy the particles and detect potential threats, macrophages express an array of receptors to sense and phagocytose prey particles. In this study, we accurately quantified a proteomic time-course of isolated phagosomes from murine bone ...[more]

PMID: 25755298

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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 Âµm 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

Dataset Information

Temporal and ligand effects on phagosome maturation

Publications

Quantitative proteome analysis of temporally resolved phagosomes following uptake via key phagocytic receptors.

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