Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse macrophages infected with B. melitensis

ABSTRACT: Facultative intracellular Brucella infect and survive inside macrophages, and the outcome of macrophage-Brucella interaction is a basis for establishment of a chronic Brucella infection. The majority of Brucella are killed at the early infection stage. A subpopulation of virulent Brucella strains is instead trafficked to an intracellular replicative phagosome, and are resistant to further attack and begin to multiply dramatically. Virulent Brucella also inhibit macrophage apoptosis that in turn favors pathogen survival and replication. We used the Affymetrix mouse GeneChip 430 2.0 array to analyze mouse macrophage gene expression profiles during the time course of virulent B. melitensis strain 16M infection. Experiment Overall Design: Murine J774.A1 macrophage cells were infected with B. melitensis strain 16M at a MOI of 200:1. Brucella cultures derived from different Brucella colonies were used to infect different groups of macrophage cells to reflect independent infections. Following 4 h incubation, the cells were washed with PBS and treated with 50 ug/ml gentamicin to kill extracellular Brucella. At 0 h (no Brucella infection), 4 h, 24 h, and 48 h post-infection, cells were individually collected, and total RNA was isolated. The Affymetrix mouse GeneChip 430 2.0 array was used for microarray hybridization. Microarray intensity data were obtained by using Affymetrix GCOS software and further analyzed by GeneSpring and other software programs.

ORGANISM(S): Mus musculus

SUBMITTER: Yongqun He

PROVIDER: E-GEOD-5202 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Brucella melitensis triggers time-dependent modulation of apoptosis and down-regulation of mitochondrion-associated gene expression in mouse macrophages.

He Yongqun Y Reichow Sherry S Ramamoorthy Sheela S Ding Xicheng X Lathigra Raju R Craig Johanna C JC Sobral Bruno W S BW Schurig Gerhardt G GG Sriranganathan Nammalwar N Boyle Stephen M SM

Infection and immunity 20060901 9

Brucella spp. are facultative intracellular bacteria that cause brucellosis in humans and other animals. Brucella spp. are taken up by macrophages, and the outcome of the macrophage-Brucella interaction is a basis for establishment of a chronic Brucella infection. Microarrays were used to analyze the transcriptional response of the murine macrophage-like J774.A1 cell line to infection with virulent Brucella melitensis strain 16M. It was found that most significant changes in macrophage gene tran ...[more]

PMID: 16926395

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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

Transcription profiling of mouse macrophages infected with B. melitensis

Publications

Brucella melitensis triggers time-dependent modulation of apoptosis and down-regulation of mitochondrion-associated gene expression in mouse macrophages.

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