Project description:RNA samples from chicken (Gallus gallus) embryonic fibroblasts (sample ID: ES90) as well as from the HD11 chicken macrophage cell line (sample ID: ES91) were sequenced at the National Institutes of Health Intramural Sequencing Center (NISC) using the Illumina GAIIx technology running the standard CASAVA/ELAND pipeline.
Project description:HD11 cells were stimulated with 1 ug/ml endotoxin from ST-798 for 1, 2, 4 and 8 hours Cells were harvested at 1, 2, 4 and 8 hrs post stimulation and RNA was isolated from these cells and microarray was conducted for these RNA samples using affymetrix genechips
Project description:We infected HD11 macrophages with Salmonella Typhimurium, followed by chemical proteomic analysis of deubiquitinases by using ubiquitin-specific active-site probe.
Project description:HD11 cells were stimulated with 1 ug/ml endotoxin from ST-798 for 1, 2, 4 and 8 hours Cells were harvested at 1, 2, 4 and 8 hrs post stimulation and RNA was isolated from these cells and microarray was conducted for these RNA samples using affymetrix genechips The microarray experiment was conducted using three replications. The first two replications each used one experimental unit and one Affymetrix GeneChip for each of the eight combinations of endotoxin dose (treated vs. control) and time (1, 2, 4, or 8 hours after treatment). The third replication was analyzed with four GeneChips for four endotoxin-treated experimental units measured at 1, 2, 4, and 8 hours after treatment, respectively.
Project description:Salmonella enterica is one of the most important foodborne pathogens that infect a variety of animals and birds. In humans, S. Typhimurium causes gastroenteritis, leading to vomiting, diarrhea, fever, and abdominal cramps. We mainly get infected with Salmonella by ingesting comminated poultry products. Therefore, developing an oral live attenuated vaccine for the poultry industry is our best bet against Salmonella infection. In this article, we investigated the potential of the next generation of Salmonella vaccines. We generated a library of potentially attenuated S. Typhimurium mutants and compared fitness to that of a commercial vaccine. We also investigated the invasion and survival potential of these mutants in chicken macrophages. Our data indicate that although these mutants had no significant growth defects, they were much sensitive to macrophage attack. Analyzing the transcriptome data from infected primary chicken macrophages, we concluded that these mutants elicit a robust immune response by activating several immunoregulatory pathways. Our data also indicates that by combining phoPQ deletion with an already existing cya-crp deletion in MeganVac1, a much stronger immune response can be generated.
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.
Project description:We present genome-wide transcript expression profiles of Salmonella Typhimurium 14028s under both macrophage in vitro and in vivo conditions. These transcriptome data were integrated with ChIP-mini datasets to elucidate the transcriptional roles of H-NS and RpoD under macrophage in vivo conditions.
