Abstract
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.
Highlights
Non-typhoid Salmonella enterica serovars such as Salmonella Enteritidis
Chicken embryo fibroblast (CEF) cell cultures were purchased from the Virology Laboratory, Veterinary Diagnostic Directorate of the National Food Chain Safety Office of Hungary, where freshly prepared cultures of chicken embryo fibroblasts (CEFs) are routinely used for virus isolation
None of the genes inducible in CEFs coded for effector proteins involved in pathogen inactivation such as lysozyme, ExFABP or other antimicrobial peptides, e.g. cathelicidins or gallinacins [7]
Summary
Non-typhoid Salmonella enterica serovars such as Salmonella Enteritidis (S. Enteritidis) are some of the most important pathogens causing gastroenteritis in humans. Chicken fibroblasts are widely used for the studying the interactions between chickens and avian viruses [12, 13], but their role in the response to Salmonella has been poorly characterised [14]. Typhimurium by the induction of TLR15 expression [14] All of these findings have indicated that fibroblasts participate in the interaction between Salmonella and the host. Their response to Salmonella infection has never been characterised in genome-wide studies. This is why we were interested whether fibroblasts respond to S. Enteritidis infection and to what extent their response differs from that of other cells and caecal tissue. G0S2 protein is involved in the control of lipid availability for β-oxidation of fatty acids in mitochondria, which may affect production of reactive oxygen species during inflammatory response
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