Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome profiling of Listeria monocytogenes during growth on a RTE-meat matrix

ABSTRACT: The gene expression profiles of L. monocytogenes strain F2365 grown in BHI agar (reference) or on a turkey deli meat (query) was examined. Aminosilane-coated cDNA microarray slide was used. The data from the normalized ratio of query to reference signal for each spot were transformed using a log2 scale. The expression level of genes obtained from the mean of each gene on the twelve slides was considered significantly different over 1.5 fold changes. Thirty nine and forty five genes in L. monocytogenes grown on a turkey meat matrix were upregulated and downregulated, respectively. To examine the gene expression profiles of L. monocytogenes grown in BHI agar or on a turkey deli meat, L. monocytogenes strains 4b F2365 aminosilane-coated cDNA microarray slide was used. Cy3 and Cy5 dyes were used for cDNA labelling. Data were obtained from three biological and four technical replicates (n = 12). Each biological replicate included a flip dye assay. TM4 package developed by TIGR was used to analyze the data.

ORGANISM(S): Listeria monocytogenes

SUBMITTER: Dongryeoul Bae

PROVIDER: E-GEOD-20274 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Data presented here show that gene expression in the mouse brain changes during systemic infection relative to a steady state (uninfected control) mouse during systemic Listeria monocytogenes infection. Microarray analysis of gene expression in the brain following i.v. inoculation of 1-2 LD 50 L. monocytogenes showed changes in brain transcriptional activity occurred prior to brain infection. Keywords: infectious diseases model, gene expression profiling for cell signaling/trafficking Mice were injected with 1-2 LD50 wild type L. monocytogenes or with sterile PBS (control) then seven animals from each group were harvested daily for 4 days. From each day, groups of five infected animals were selected for microarray analysis based upon similarities in CFU bacteria in the spleen (day 1), blood (day 2), and brain (days 3 and 4) and were randomly paired with brains harvested from control animals. RNA was extracted from the brain halves using the Atlasâ¢ Pure Total RNA Labeling System and hybridized to plastic slides with 32P-labeled probes using Atlasâ¢ Plastic Mouse 5K Microarray (Clontech, Mountain View, CA) according to the manufacturerâs directions. Hybridization to the arrays and phosphorimaging analysis were performed in the OUHSC microarray facility. Phosphorimaging analysis was performed using a Stormâ¢ optical scanner (Molecular Dynamics Inc., Sunnyvale, CA), and initial background corrections were performed using the Array Vision Software (Imaging Research Inc., St. Catharines, Ontario). Data analyses were performed using Genespring GX v. 7.3 (Agilent Technologies, Palo Alto, CA). Raw signal data were normalized to the median of the entire chip for each sample, and all samples were further normalized per gene to the medians of data from the uninfected samples. Data for each day were analyzed by Mann Whitney Wilcoxen nonparametric test with a p-value cutoff of 0.05. Lists of genes were generated to reflect 2, 3, or 5-fold upregulation and downregulation based on significance. Expression data were incorporated into Ingenuity software (Ingenuity Systems, Inc., Redwood City, CA) to enable further understanding of complex relationships as well as provide further information about key canonical pathways in which the genes of interest functions.

Project description:Objective: Reg3g has been proposed to have a protective role against infection due to its bactericidal effect on Gram-positive bacteria, but evidence from in vivo studies is lacking. Therefore we generated a Reg3g-/- mouse, to determine its role in intestinal homeostasis and protection against experimental infection. Methods: Reg3g-/- mice were phenotyped using histological methods and a range of innate and immune markers. To investigate the antimicrobial role of Reg3g we experimentally infected mice with Gram-positive Listeria monocytogenes and Gram-negative Salmonella entertitidis and measured translocated bacteria, mucosal and systemic markers of infection. Results: Reg3g-/- mice display altered ileal mucus distribution and increased bacterial contact with the epithelium. , concomitant with This increased the inflammatory status in of the ileal mucosa and increased expression of Il-22, myeloperoxidase (MPO) and serum chemokines in serum. In response to infection, Reg3g-/- mice showed transcriptome changes and elevated levels of mucosal MPO in the ileum, but no increased bacterial translocation to the organs. Conclusions: Reg3g is equally distributed throughout the mucus of wild type (wt) mice and its absence results in an altered distribution of the ileal mucus. Reg3g deficiency also results resulted in increased bacterial contact with the epithelium and heightened inflammatory responses in the ileal mucosa. We propose that Reg3g binds pathogens suggesting it and contributes to mucus barrier function by ensnaring bacteria. Compared to wt mice, Reg3g-/- mice infected with S. enteritidis and L. monocytogenes show an increase of mucosal inflammatory markers indicating the protective, anti-microbial roles of Reg3g in defence against both Gram-positive and -negative bacteria. This study was set up according to a one-treatment, one-control design; treatments were inoculation with either Listeria monocytogenes or Salmonella enteritidis bacterial pathogens. The study results contain transcriptional profiles from infected and sham-infected control C57Bl/6 mice. In total, this study includes data from 2 treatments and 1 control of (pooled) wild-type C57Bl/6 mice and Reg3g-/- KO mutant C57Bl/6 mice = 6 arrays.

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Transcriptome profiling of Listeria monocytogenes during growth on a RTE-meat matrix

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