Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse Peyers patches after infection of C57BL/6 strain with Yersinia enterocolitica

ABSTRACT: Orogastral infection of mice with Yersinia enterocolitical leads to HIF-1 alpha activation.To elucidate whether this HIF-1 alpha activation also results in a HIF-1 dependent gene programming, the transcriptomes from Peyers Patches of uninfected and Yersinia enterocolitica infected mice were analyzed by means of of microarray analyses using Affymetrix GeneChip probe arrays (MG-U74Av2). In total, 288 genes were differentially regulated three day after infection in PP compared with the expression of uninfected control mice. Of these 288 genes, 217 were found to be differentially upregulated and from these, 14 genes ( 6.5% of all upregulated genes) are well described to be regulated via HIF-1. These data indicate that orogatral infection with Y. enterocolitica results in HIF-1 dependent gene programmning Experiment Overall Design: Per group five C57BL/6 mice were infected orogastrally with 500 Million Yersinia enterocolitica. 1 and 3 days after infection, Peyers Patches were removed and total RNA was prepared. In parallel, RNA was isolated from uninfected mice. The generation of fragmented cRNA was performed following the manufacturers instructions and used for hybridization onto GeneChip arrays MG-U74Avs2. Analysis of microarray data was performed using the Affymetrix Microarray Suite 5.0, Affymetrix Mining Tool 3.0. A median signal log2 ratio (SLR) grater than 1.5 or less than -1.5 was considered a significant change.

ORGANISM(S): Mus musculus

SUBMITTER: Erwin Bohn

PROVIDER: E-GEOD-9558 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background:; Yersinia outer protein (Yop) H is a secreted virulence factor of Yersinia enterocolitica which inhibits phagocytosis of Y. enterocolitica and promotes virulence of Y. enterocolitica (Ye) in mice. The aim of this study was to address whether and how YopH affects the innate immune response against Ye in mice. Results:; For this purpose mice were infected with wild type Ye (pYV+) or a YopH-deficient Ye mutant strain (DyopH). CD11b+ cells were isolated from infected spleen and subjected to gene expression analysis using microarrays. Despite attenuation of DyopH in vivo, by variation of infection doses we were able to achieve conditions that allow comparison of gene expression in pYV+ and DyopH infections at either comparable infection courses or splenic bacterial burden. Gene expression analysis provided evidence that expression levels of several immune response genes including IFN-g and IL-6 are high after pYV+ infection but low after sublethal DyopH infection. In line with these findings, infection of IFN-gR-/- and IL-6-/- mice with pYV+ or DyopH revealed that these cytokines are dispensable for control of DyopH, but not pYV+ infection. Consistently, in bacteria killing assays with BMM in vitro, stimulation of BMM with IFN-g is required for killing of pYV+ but not DyopH. Conclusion:; In conclusion, this data suggest that IFN-g counteracts YopH-mediated virulence mechanisms of Ye which in Ye wild type infection contribute to evasion of the innate immune response including killing by macrophages. Experiment Overall Design: In this study microarray analyses were performed to define differences in gene expression of cells associated with innate immune response (CD11b+ cells) after infection of mice with a sublethal and lethal infection with wildtype Yersinia enterocolitica compared to uninfected mice. In addition, we wanted to investigate whether differences in gene expression can be defined which are due to the virulence factor YopH. Moroeover, we were interested whether gene expression pattern of sublethal and lethal infected mice are different. Allover all five samples were compared. Number of replicates 1.

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Transcription profiling of mouse Peyers patches after infection of C57BL/6 strain with Yersinia enterocolitica

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