Unknown,Transcriptomics,Genomics,Proteomics

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Kinetics of host transcription in CEFs productively infected with recombinant fowlpox virus vaccine vector FP9

ABSTRACT: Fowlpox virus (here as FP9, a plaque-purified, high passage-attenuated derivative) effectively suppresses induction of the âinnateâ immune responses, notably the Type I interferon system, of the permissive host (chicken). Despite the extensive usage of fowlpox virus as a recombinant vaccine vector in chickens, its immunomodulatory mechanisms remain largely unknown. In this study, a transcriptomic analysis using the Affymetrix GeneChip chicken genome array was performed at the host gene transcription level at 4, 8, 16 and 24 hours post-infection of mock treated and FP9-infected (MOI=5, 2h) chicken embryo fibroblasts (CEF). The experiment was performed in triplicate with three different batches of CEFs. Because fowlpox virus is capable of expressing antigens in mammalian cells, these studies in chicken cells form a baseline for subsequent study of immunomodulation of mammalian innate immune responses.

ORGANISM(S): Gallus gallus

SUBMITTER: Michael Skinner

PROVIDER: E-MTAB-5455 | 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.

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Kinetics of host transcription in CEFs productively infected with recombinant fowlpox virus vaccine vector FP9

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