Unknown,Transcriptomics,Genomics,Proteomics

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Impacts of Salmonella enteritidis Infection on Liver Transcriptome in Broilers

ABSTRACT: To provide insight into the systemic metabolic effects of S. enteritidis infection, liver samples were harvested 10 days post infection from broiler hens. Hepatic global gene expression levels were assessed using a chicken 44K Agilent microarray. Forty-four genes were differentially expressed at a significance level of q-value < 0.05. One hundred eighty-three genes were differentially expressed at a suggestive significance level of q-value < 0.1. A predominance of down-regulation existed among significantly differentially expressed genes. A cell morphology, cell cycle, organismal injury and abnormalities network and a metabolic disease, cardiovascular system development and function, and urological disease network were created from the differentially expressed genes. Apoptosis, electron transport, peptidase activity, vein constriction, cell differentiation, IL-2 signaling, Jak-Stat signaling, B-cell receptor signaling, GDP/GTP exchange, and cytokine suppression were among the functions of the differentially expressed genes that were down-regulated in response to S. enteritidis. The effects of S. enteritidis infection on the liver transcriptome profiles of broilers reflect a predominance of down-regulation of genes involved with metabolic, cell morphology, cell cycle, urological disease, and organismal injury functions. These results provide insight into important systemic metabolic mechanisms that are active in the chicken liver in response to S. enteritidis infection. Sixteen broiler hens of approximately 5 months (8 challenged orally with 1x108 S. enteritidis on each of three sequential days and 8 mock-challenged with phosphate buffered saline) were used. Livers were harvested at 10 days post infection and stored in RNALater (Ambion).

ORGANISM(S): Gallus gallus

SUBMITTER: Derrick Coble

PROVIDER: E-GEOD-39448 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria. Increased levels of G0S2 might decrease the availability of fatty acids to mitochondria. In non-professional phagocytes such as CEFs, this may lead to the dysfunction of mitochondria, apoptosis of CEFs and release of intracellular Salmonella. In professional phagocytes, G0S2 might be involved in the control of mitochondrial respiration, resulting in a decrease of reactive oxygen species as respiration by-products and lower damage to tissue. In this study we were interested whether chicken embryo fibroblast (CEFs) respond to S. Enteritidis infection and to what extent their response differs from that of other cells and caecal tissue. To address this, we characterised the gene expression of CEFs after infection with two different wild-type S. Enteritidis strains of poultry origin - SE 147 and SE 11 - using Agilen custom 8×15K microarrays. In total, 13,681 probes were designed to characterise the expression of ~9,000 transcripts of Gallus gallus.

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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Impacts of Salmonella enteritidis Infection on Liver Transcriptome in Broilers

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