Global transcriptional response of porcine intestinal epithelial cell lines to Salmonella enterica serovar Typhimurium and Choleraesuis
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus scrofa
SUBMITTER: Rocio Bautista
PROVIDER: E-GEOD-23063 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
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 37ºC. 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 µg/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
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 37ºC. 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 µg/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
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 37ºC. 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 µg/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
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 37ºC. 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 µg/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
Project description:Mycobacterium smegmatis is a model non-pathogenic mycobacterium that is efficiently killed by macrophages. Here, we explore the role of NF-?B in the innate immune response, focusing in detail on the mechanisms of the first killing period (1-4h) of M. smegmatis which coincides with phagosome-lysosome fusion. We show that infection of macrophages with M. smegmatis induces an activation of NF-?B and this activation is required for killing since treatment of macrophages with NF-?B inhibitors or siRNA silencing of the NF-?B subunit p65 increases bacterial survival. NF-?B induced proteins were thus hypothesized to be essential during the first phase of M. smegmatis killing. We therefore identified, using RNA microarray, the genes that were regulated during infection in the absence and presence of NF-?B inhibitors. By subtraction this provided a list of pro-inflammatory proteins that were under the control of NF-?B and putatively involved in the killing response. Among these category of genes were those for lysosomal enzymes and membrane trafficking regulators, including Cathepsins, LAMP-2 and Rab34, are regulated by NF-?B. Moreover, inhibition of NF-?B signaling retarded the delivery of v-ATPase, LAMP-2, CtsZ and CtsH thereby impairing the maturation of mycobacterial phagosomes. Collectively; our data provide the first compelling evidence that the innate immune response via NF-?B activation is linked to phagosome fusion with lysosomes that is essential for killing of mycobacteria. Keywords: NFkB inhibitor treated vs untreated The study includes J774 macrophage cell lines which are infected with Mycobacterium smegmatis in the presence and absense of NFkB inhibitor SC-514.Total RNA was isolated from individual samples and each sample was hybridised to CodeLink Mouse whole genome bioarray slides.This study was intended to know the role of NFkB regulated genes in killing non-pathogenic mycobacteria during 4 hour post infection of macrophages.
2007-10-23 | E-GEOD-8999 | biostudies-arrayexpress
Project description:Analysis of the human monocyte-derived macrophage (hMDM) transcriptional response to L. pneumophila infection at 8 hours post-infection We used microarrays to probe the hMDM transcriptional response to wild type L. pneumophila AA100 infection and its isogenic ankB and T2SS mutants. We identified up-regulation of innate immunity pathways and down-regulation of protein translation pathways hMDMs were isolated from healthy donors and infected with L. pneumophila AA100 and its isogenic ankB and T2SS mutants for 8 h prior to isolation of total RNA and hybridization to Affymetrix microarrays.
2014-09-19 | E-GEOD-61535 | biostudies-arrayexpress
Project description:The innate immune system provides the first response to pathogen infection and orchestrates the activation of the adaptive immune system. Though a large component of the innate immune response is common to all infections, pathogen-specific innate immune responses have been documented as well. The innate immune response is thought to be especially critical for fighting infection with Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB). While TB can be a deadly disease, only 5-10% of individuals infected with MTB develop active disease, and this inter-individual variation is, at least partly, heritable. Studies of inter-individual variation in the innate immune response to MTB infection may therefore shed light on the genetic basis for variation in susceptibility to TB. Yet, to date, we still do not know which properties of the innate immune response are specific to MTB infection and which represent a general response to pathogen infection. To begin addressing this gap, we infected macrophages with eight different bacterial pathogens, including different MTB strains and related mycobacteria, and studied the transcriptional response to infection. We found that although the gene expression changes were largely consistent across the bacterial infection treatments, we were able to identify a novel subset of genes whose regulation was affected specifically by infection with mycobacteria. Genetic variants that are associated with regulatory differences in these genes should be considered candidate loci for explaining inter-individual susceptibility TB. RNA-seq of monocyte-derived macrophages isolated from 6 healthy European males at 4, 18, and 48 hours post-infection with the following 8 bacteria: Mycobacterium tuberculosis (MTB) H37Rv, Mycobacterium tuberculosis GC1237, MTB GC1237, bacillus Calmette-Guérin (BCG), Mycobacterium smegmatis, Yersinia pseudotuberculosis, Salmonella typhimurium, and Staphylococcus epidermidis. table-s1.txt is a tab-delimited text file that contains the batch-corrected log2 counts per million for each of the 156 samples, as well as the Ensembl gene ID and gene name. BCG = bacillus Calmette-Guérin GC = Mycobacterium tuberculosis GC1237 Rv = Mycobacterium tuberculosis (MTB) H37Rv Rv+ = heat-inactivated MTB H37Rv Salm = Salmonella typhimurium Smeg = Mycobacterium smegmatis Staph = Staphylococcus epidermidis Yers = Yersinia pseudotuberculosis
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