Project description:The GeneChip Porcine Genome Array was used to identify the transcriptional response upon either Salmonella typhimurium (ST) or Salmonella choleraesuis (SC) infection in two porcine epithelial cell lines (IPEC-J2, from jejunum and IPI-2I, from ileum) during 2 and 4 hours post infection. The objectives in this study were first, to identify the different response between the epithelial cell lines from different gut regions; second, to study how the Salmonella serotypes used could elicit a different host response; and third, to determine the effect of the time-points on the differentially gene expression. Epithelial cells were seeded into 6-well tissue culture plates and grown to confluence in 5% CO2 at 37ÂºC. Monolayers were infected for 1 h. with Salmonella typhimurium or Salmonella choleraesuis serotypes (MOI 1:10) or incubated with media alone (Control cells). Extracellular bacteria were removed, and cultures were further incubated during 2 and 4 h. in the presence of 50 Âµg/ml of the non-cell-permeant antibiotic gentamicin to kill remaining extracellular bacteria. The experiment was developed in triplicate in order to ensure a proper robustness in the microarray statistical analysis. An indirect ELISA was used to confirm cell activation by means of IL8 detection in cell culture medium. After each time-point, cells were lysed and RNA extracted.

2013-12-20 | E-GEOD-23057 | biostudies-arrayexpress

Global transcriptional response of porcine intestinal epithelial cell lines to Salmonella enterica serovar Typhimurium and Choleraesuis: IPI-2I infected with S. choleraesuis

Project description:The GeneChip Porcine Genome Array was used to identify the transcriptional response upon either Salmonella typhimurium (ST) or Salmonella choleraesuis (SC) infection in two porcine epithelial cell lines (IPEC-J2, from jejunum and IPI-2I, from ileum) during 2 and 4 hours post infection. The objectives in this study were first, to identify the different response between the epithelial cell lines from different gut regions; second, to study how the Salmonella serotypes used could elicit a different host response; and third, to determine the effect of the time-points on the differentially gene expression. Epithelial cells were seeded into 6-well tissue culture plates and grown to confluence in 5% CO2 at 37ÂºC. Monolayers were infected for 1 h. with Salmonella typhimurium or Salmonella choleraesuis serotypes (MOI 1:10) or incubated with media alone (Control cells). Extracellular bacteria were removed, and cultures were further incubated during 2 and 4 h. in the presence of 50 Âµg/ml of the non-cell-permeant antibiotic gentamicin to kill remaining extracellular bacteria. The experiment was developed by triplicate in order to ensure a proper robustness in the microarray statistical analysis. An indirect ELISA was used to confirm cell activation by mean of IL8 detection in cell culture medium. After each time-point, cells were lysed and RNA extracted.

2013-12-20 | E-GEOD-23060 | biostudies-arrayexpress

Global transcriptional response of porcine intestinal epithelial cell lines to Salmonella enterica serovar Typhimurium and Choleraesuis: IPI-2I infected with S. typhimurium

Project description:The GeneChip Porcine Genome Array was used to identify the transcriptional response upon either Salmonella typhimurium (ST) or Salmonella choleraesuis (SC) infection in two porcine epithelial cell lines (IPEC-J2, from jejunum and IPI-2I, from ileum) during 2 and 4 hours post infection. The objectives in this study were first, to identify the different response between the epithelial cell lines from different gut regions; second, to study how the Salmonella serotypes used could elicit a different host response; and third, to determine the effect of the time-points on the differentially gene expression. Epithelial cells were seeded into 6-well tissue culture plates and grown to confluence in 5% CO2 at 37ÂºC. Monolayers were infected for 1 h. with Salmonella typhimurium or Salmonella choleraesuis serotypes (MOI 1:10) or incubated with media alone (Control cells). Extracellular bacteria were removed, and cultures were further incubated during 2 and 4 h. in the presence of 50 Âµg/ml of the non-cell-permeant antibiotic gentamicin to kill remaining extracellular bacteria. The experiment was developed by triplicate in order to ensure a proper robustness in the microarray statistical analysis. An indirect ELISA was used to confirm cell activation by mean of IL8 detection in cell culture medium. After each time-point, cells were lysed and RNA extracted.

2013-12-20 | E-GEOD-23062 | biostudies-arrayexpress

Global transcriptional response of porcine intestinal epithelial cell lines to Salmonella enterica serovar Typhimurium and Choleraesuis: IPEC-J2 infected with S. typhimurium

Project description:The GeneChip Porcine Genome Array was used to identify the transcriptional response upon either Salmonella typhimurium (ST) or Salmonella choleraesuis (SC) infection in two porcine epithelial cell lines (IPEC-J2, from jejunum and IPI-2I, from ileum) during 2 and 4 hours post infection. The objectives in this study were first, to identify the different response between the epithelial cell lines from different gut regions; second, to study how the Salmonella serotypes used could elicit a different host response; and third, to determine the effect of the time-points on the differentially gene expression. Epithelial cells were seeded into 6-well tissue culture plates and grown to confluence in 5% CO2 at 37ÂºC. Monolayers were infected for 1 h. with Salmonella typhimurium or Salmonella choleraesuis serotypes (MOI 1:10) or incubated with media alone (Control cells). Extracellular bacteria were removed, and cultures were further incubated during 2 and 4 h. in the presence of 50 Âµg/ml of the non-cell-permeant antibiotic gentamicin to kill remaining extracellular bacteria. The experiment was developed by triplicate in order to ensure a proper robustness in the microarray statistical analysis. An indirect ELISA was used to confirm cell activation by mean of IL8 detection in cell culture medium. After each time-point, cells were lysed and RNA extracted.

2013-12-20 | E-GEOD-23059 | biostudies-arrayexpress

Global transcriptional analysis of dehydrated Salmonella enterica serovar Typhimurium.

Project description:Despite the scientific and industrial importance of desiccation tolerance in Salmonella, knowledge regarding its genetic basis is still scarce. In the present study, we performed a transcriptomic analysis of dehydrated and water-suspended Salmonella enterica serovar Typhimurium using microarrays. Dehydration induced expression of 90 genes and downregulated that of 7 genes. Ribosomal structural genes represented the most abundant functional group with a relatively higher transcription during dehydration. Other main induced functional groups included genes involved in amino acid metabolism, energy production, ion transport, transcription, and stress response. The highest induction was observed in the kdpFABC operon, encoding a potassium transport channel. Knockout mutations were generated in nine upregulated genes. Five mutants displayed lower tolerance to desiccation, implying the involvement of the corresponding genes in the adaptation of Salmonella to desiccation. These included genes encoding the isocitrate-lyase AceA, the lipid A biosynthesis palmitoleoyl-acyltransferase Ddg, the modular iron-sulfur cluster scaffolding protein NifU, the global regulator Fnr, and the alternative sigma factor RpoE. Notably, these proteins were previously implicated in the response of Salmonella to oxidative stress, heat shock, and cold shock. A strain with a mutation in the structural gene kdpA had a tolerance to dehydration comparable to that of the parent strain, implying that potassium transport through this system is dispensable for early adaptation to the dry environment. Nevertheless, this mutant was significantly impaired in long-term persistence during cold storage. Our findings indicate the involvement of a relatively small fraction of the Salmonella genome in transcriptional adjustment from water to dehydration, with a high prevalence of genes belonging to the protein biosynthesis machinery.

| S-EPMC3485933 | biostudies-literature

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