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Candida albicans phagocytosed by macrophages: intact vs. ruptured phagosomes

ABSTRACT: Transcriptional profiling of Candida albicans after 3 h phagocytosis by vehicle DMSO-treated macrophages (intact, expanding phagosomes) or calcium chelator BAPTA-AM-treated macrophages (inhibits lysosomal repair of expanding phagosomes, leading to phagosome rupture) to determine the effect of preventing phagosome expansion on C. albicans gene expression after phagocytosis by macrophages. Cultivation of Candida only for 3 h in DMEM-FBS cell culture medium or YPD complex medium as non-phagocytosis control conditions.

ORGANISM(S): Candida albicans

PROVIDER: GSE147207 | GEO | 2020/10/05

REPOSITORIES: GEO

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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

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Candida albicans phagocytosed by macrophages: intact vs. ruptured phagosomes

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