Characterization of the in vitro gene response of chicken cells to Salmonella Enteritidis

Szmolka, Ama and Matulova, Marta and Wiener, Zoltán and Nagy, Béla and Rychlik, Ivan (2014) Characterization of the in vitro gene response of chicken cells to Salmonella Enteritidis. In: EMBO Conference on Microbiology after the genomics revolution: Genomes 2014, 24–27 June 2014, Paris.

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Abstract

Salmonella Enteritidis (SE) is one of the most frequently reported causative agent of human gastroenteritis, originating mainly from poultry. Pathogenesis of SE infection in poultry is well-elucidated, but the complexity of the host cell response, and its relation to differring pathogenic potential of various strains is much less understood. Therefore we intended to provide a genome-wide comparative characterization of the gene expression profiles of chicken cells to wild type strains and virulence-related mutants of Salmonella Enteritidis. Freshly isolated chicken embryo fibroblast (CEF) cells co-incubated with Salmonella for 4 hrs were used to model gene response of young chickens to Salmonella infection and to measure the invasiveness of wild type strains SE147, SE11 and non-motile mutants of SE11 lacking the fliD gene and/or the virulence plasmid. Agilent custom 8×15K microarray was designed to profile the expression of 13741 chicken genes, with emphasis to those related to immune response. Significant gene expression changes with fold change ≥3 (in total of 31 genes) were verified by real-time PCR. Expression profile of infected CEF cells resulted in 314 genes significantly misregulated by the infection with the wild type strain SE147 (206 up-/108 down-regulations) while only 135 genes were significantly expressed as a result to SE11 infection (74 up-/61 down-regulations). There were 100 genes induced by both wild strains, among them CSF3 (colony-stimulating factor), IL-1β and IL-8 showing the highest upregulations. In contrast to this, infection with non-motile mutants lacking fliD gene and/or the virulence plasmid, did not cause any significant change in host gene expression. However real-time PCR results indicated that the cell cycle-related G0S2 switch-, and the enolase ENO2 genes were highly induced by the mutant strains, indicating that the reduced invasiveness of the mutants might have stimulated cell division and/or metabolism of the host cells. Results suggest that fliD gene plays a key role in the invasiveness of Salmonella strains, and could be considered as an important modulator of the chicken response to Salmonella infection. This work was supported by the EU FP6 NoE MedVetNet and OTKA 105635. Ama Szmolka is a holder of János Bolyai Research Scholarship of HAS.

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