Transcriptional Characterization of Streptococcus pneumoniae Interacting with Human Pharyngeal Cells
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ABSTRACT:
ORGANISM(S): Streptococcus pneumoniae TIGR4
SUBMITTER: Jerry Liu
PROVIDER: E-GEOD-44947 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:We used a combination of adherence assays, mutagenesis and functional genomics to identify novel factors involved in adherence. This work identifies a list of novel potential pneumococcal adherence determinants.
2013-09-30 | GSE44947 | GEO
Project description:Infection of the human host by Streptococcus pneumoniae begins with colonization of the nasopharynx, which is mediated by adherence of bacteria to respiratory epithelium. Several studies have indicated an important role for the pneumococcal capsule in this process. Here, we used microarrays to characterize the in vitro transcriptional response of human nasopharyngeal epithelial Detroit 562 cells to adherence of serotype 2-encapsulated strain D39, serotype 19F-encapsulated strain G54, serotype 4-encapsulated strain TIGR4, and their nonencapsulated derivatives (delta-cps). In total, 322 genes were found to be upregulated in response to adherent pneumococci. Twenty-two genes were commonly induced, including those encoding several cytokines (e.g., IL-1-beta, IL-6), chemokines (e.g., IL-8, CXCL1/2), and transcriptional regulators (e.g., FOS), consistent with an innate immune response mediated by Toll-like receptor signaling. Interestingly, 85% of genes was induced specifically by one or more encapsulated strains, suggestive of a capsule-dependent response. Importantly, purified capsular polysaccharides alone had no effect. Over a third of these loci encoded products predicted to be involved in transcriptional regulation and signal transduction, in particular MAPK signaling pathways. Real-time PCR of a subset of ten genes confirmed microarray data and showed a time-dependent upregulation of especially innate immunity genes. Downregulation of epithelial genes was most pronounced upon adherent D39delta-cps, as 68% of the 161 genes identified was only repressed using this nonencapsulated strain. In conclusion, we identified a subset of host genes specifically induced by encapsulated strains during in vitro adherence, and have demonstrated the complexity of interactions occurring during the initial stages of pneumococcal infection. Experiment Overall Design: We used three different pneumococcal strains and their isogenic nonencapsulated derivatives (delta-cps): serotype 2-encapsulated strain D39, serotype 19F-encapsulated strain G54, and serotype 4-encapsulated strain TIGR4. All experiments were performed in triplicate (3 independent biological replicates) and compared to uninfected control Detroit 562 cells. Bacteria were allowed to adhere to the epithelial cells for 2 hours, after which the transcriptional response of the Detroit cells was analyzed using Affymetrix Human U133 Plus GeneChips. In addition to the 6 strains mentioned above, we included transcriptional analysis of the epithelial cell response to low-dose D39delta-cps (giving adherence equivalent to wild-type D39) and purified type 2 capsular polysaccharides.
2008-06-16 | E-GEOD-8527 | biostudies-arrayexpress
Project description:Infection of the human host by Streptococcus pneumoniae begins with colonization of the nasopharynx, which is mediated by adherence of bacteria to respiratory epithelium. Several studies have indicated an important role for the pneumococcal capsule in this process. Here, we used microarrays to characterize the in vitro transcriptional response of human nasopharyngeal epithelial Detroit 562 cells to adherence of serotype 2-encapsulated strain D39, serotype 19F-encapsulated strain G54, serotype 4-encapsulated strain TIGR4, and their nonencapsulated derivatives (Δcps). In total, 322 genes were found to be upregulated in response to adherent pneumococci. Twenty-two genes were commonly induced, including those encoding several cytokines (e.g., IL-1β, IL-6), chemokines (e.g., IL-8, CXCL1/2), and transcriptional regulators (e.g., FOS), consistent with an innate immune response mediated by Toll-like receptor signaling. Interestingly, 85% of genes was induced specifically by one or more encapsulated strains, suggestive of a capsule-dependent response. Importantly, purified capsular polysaccharides alone had no effect. Over a third of these loci encoded products predicted to be involved in transcriptional regulation and signal transduction, in particular MAPK signaling pathways. Real-time PCR of a subset of ten genes confirmed microarray data and showed a time-dependent upregulation of especially innate immunity genes. Downregulation of epithelial genes was most pronounced upon adherent D39Δcps, as 68% of the 161 genes identified was only repressed using this nonencapsulated strain. In conclusion, we identified a subset of host genes specifically induced by encapsulated strains during in vitro adherence, and have demonstrated the complexity of interactions occurring during the initial stages of pneumococcal infection. Keywords: Paired measurements
2007-10-01 | GSE8527 | GEO
Project description:The polyamine transport operon in Streptococcus pneumoniae TIGR4 is necessary for survival in murine models of pneumococcal pneumonia. To date, there is no description of polyamine transport dependent pneumococcal gene expression. In this study, we compared gene expression between the wild-type and transport deficient (potABCD) TIGR4 by RNA-Seq analysis.
2017-08-14 | E-MTAB-5951 | biostudies-arrayexpress
Project description:The polyamine biosynthesis gene, speE, in Streptococcus pneumoniae TIGR4 is necessary for survival in murine models of pneumococcal pneumonia. To date, there is no description of polyamine biosynthesis dependent pneumococcal gene expression. In this study, we compared gene expression between the wild-type and biosynthesis deficient (speE) TIGR4 by RNA-Seq analysis.
2023-01-01 | E-MTAB-6070 | biostudies-arrayexpress
Project description:The aim of this experiment is to determine the difference in protein expression, if any, in a mouse model of pneumococcal pneumonia using wild type and a polyamine transport operon deletion mutant strain of S. pneumoniae TIGR4, using expression proteomics. Eight week old female C57bl/6 mice were used in this study. The experimental design had two groups of 3 animals each that were administered 50 µL 1 X 107 CFU wild type S. pneumoniae TIGR4 or S. pneumoniae TIGR4 ΔpotABCD strain by intranasal instillation. The control group with three animals received 50 µL PBS. Mice were euthanized 4 hr and 12 hr post infection and lung tissues were harvested. The control group animals were euthanized 12 hr post infection.
2018-10-27 | PXD002300 | Pride
Project description:Recent murine studies have demonstrated that the role of response regulator 09 (RR09) of S. pneumoniae in virulence varies between strains. In the present study, we used a murine pneumonia model of infection to assess the virulence of a TIGR4 rr09-mutant, and found that TIGR4Δrr09 was attenuated after intranasal infection. Further, we investigated the in vitro transcriptional changes in pneumococcal rr09 mutants of two strains, D39 and TIGR4, by microarray analysis. The transcriptional profiles of the rr09 mutants in both strains displayed clear differences compared to their parental wild-type. In D39Δrr09 but not TIGR4Δrr09, genes involved in competence (e.g. comAB) were upregulated. In TIGR4, genes located on the rlrA pathogenicity islet, absent in the D39 genome, appeared to be regulated by RR09. Furthermore, several phosphotransferase systems (PTSs), believed to be involved in sugar uptake (e.g., sp0060-sp0066), were strongly downregulated in D39Δrr09, while not regulated by RR09 in TIGR4. To address the role of one of these PTS in virulence, D39Δsp0063 was constructed and tested in a murine infection model. No difference in virulence compared to the wild-type was found, indicating that downregulation of this gene alone is not the cause of the avirulent phenotype of D39Δrr09. Finally, expression of rr09 and three of our identified RR09 targets were assessed during infection in mice. This in vivo experiment confirmed differences in expression between TIGR4 wild-type and rr09-mutant, as well as between wild-type D39 and TIGR4. In conclusion, our results indicate strain-specific regulation of pneumococcal gene expression by RR09. Keywords: genetic modification
2006-12-24 | GSE6137 | GEO
Project description:Recent murine studies have demonstrated that the role of response regulator 09 (RR09) of S. pneumoniae in virulence varies between strains. In the present study, we used a murine pneumonia model of infection to assess the virulence of a TIGR4 rr09-mutant, and found that TIGR4Δrr09 was attenuated after intranasal infection. Further, we investigated the in vitro transcriptional changes in pneumococcal rr09 mutants of two strains, D39 and TIGR4, by microarray analysis. The transcriptional profiles of the rr09 mutants in both strains displayed clear differences compared to their parental wild-type. In D39Δrr09 but not TIGR4Δrr09, genes involved in competence (e.g. comAB) were upregulated. In TIGR4, genes located on the rlrA pathogenicity islet, absent in the D39 genome, appeared to be regulated by RR09. Furthermore, several phosphotransferase systems (PTSs), believed to be involved in sugar uptake (e.g., sp0060-sp0066), were strongly downregulated in D39Δrr09, while not regulated by RR09 in TIGR4. To address the role of one of these PTS in virulence, D39Δsp0063 was constructed and tested in a murine infection model. No difference in virulence compared to the wild-type was found, indicating that downregulation of this gene alone is not the cause of the avirulent phenotype of D39Δrr09. Finally, expression of rr09 and three of our identified RR09 targets were assessed during infection in mice. This in vivo experiment confirmed differences in expression between TIGR4 wild-type and rr09-mutant, as well as between wild-type D39 and TIGR4. In conclusion, our results indicate strain specific regulation of pneumococcal gene expression by RR09. Keywords: Genetic modification
2006-10-28 | GSE6139 | GEO
Project description:In the present study, we used RNA sequencing on mouse splenocytes to describe the immune recall in the days following pneumococcal lung infection. Mice were sacrificed at days 1, 2, 4, and 7 after Streptococcus pneumoniae (TIGR4 serotype 4) intranasal infection and splenocytes were cultured in the presence or absence of the same inactivated bacterial strain to access the transcriptomic profile.
2022-04-02 | GSE199605 | GEO
Project description:Previous studies have indicated that PsaR of Streptococcus pneumoniae is a manganese-dependent regulator, negatively affecting the expression of at least seven genes. Here, we extended these observations by transcriptome and proteome analysis of psaR mutants in strains D39 and TIGR4. The microarray analysis identified three shared PsaR targets: the psa-operon, pcpA, and prtA. Additionally, we found 31 genes to be regulated by PsaR in D39 only, most strikingly a cellobiose-specific PTS and a putative bacteriocin operon (sp0141-sp0146). In TIGR4, 14 PsaR gene targets were detected, with the rlrA pathogenicity islet being the most pronounced. Proteomics confirmed most of the shared gene targets. To examine the contribution of PsaR to pneumococcal virulence, we compared D39 and TIGR4 wild-type (wt) and psaR-mutants in three murine infection models. During colonization, no clear effect was observed of the psaR mutation in either D39 or TIGR4. In the pneumonia model, small but significant differences were observed in the lungs of mice infected with either D39wt or ∆psaR: D39∆psaR had an initial advantage in survival in the lungs. Conversely, TIGR4∆psaR-infected mice had significantly lower bacterial loads at 24h only. Finally, during experimental bacteremia, D39∆psaR-infected mice had significantly lower bacterial loads in the blood stream than wt-infected mice for the first 24h of infection. TIGR4∆psaR showed attenuation at 36h only. In conclusion, our results show that PsaR of D39 and TIGR4 has a strain-specific role in global gene expression and in the development of bacteremia in mice.
2008-11-19 | GSE13505 | GEO