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| Status |
Public on Oct 01, 2007 |
| Title |
Analysis of the in vitro transcriptional response of human pharyngeal epithelial cells to adherent pneumococci |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
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
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| Overall design |
We used three different pneumococcal strains and their isogenic nonencapsulated derivatives (Δcps): 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 D39Δcps (giving adherence equivalent to wild-type D39) and purified type 2 capsular polysaccharides.
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| Contributor(s) |
Bootsma HJ, Egmont-Petersen M, Hermans PW |
| Citation(s) |
17709418 |
| Submission date |
Jul 19, 2007 |
| Last update date |
Mar 25, 2019 |
| Contact name |
Hester Bootsma |
| E-mail(s) |
h.bootsma@cukz.umcn.nl
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| Phone |
+31-243666332
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| Organization name |
Radboud University Medical Centre
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| Department |
Pediatrics
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| Lab |
Laboratory of Pediatric Infectious Diseases
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| Street address |
Kapittelweg 29
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| City |
Nijmegen |
| ZIP/Postal code |
6525 EN |
| Country |
Netherlands |
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| Platforms (1) |
| GPL570 |
[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array |
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| Samples (32)
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| GSM211670 |
D39delta-cps, biological rep1 |
| GSM211671 |
D39delta-cps, biological rep2 |
| GSM211672 |
D39delta-cps, biological rep3 |
| GSM211673 |
G54delta-cps, biological rep1 |
| GSM211674 |
G54delta-cps, biological rep2 |
| GSM211675 |
G54delta-cps, biological rep3 |
| GSM211676 |
Control R1, biological rep1 |
| GSM211677 |
Control R1, biological rep2 |
| GSM211678 |
Control R1, biological rep3 |
| GSM211679 |
G54WT, biological rep1 |
| GSM211680 |
G54WT, biological rep2 |
| GSM211681 |
G54WT, biological rep3 |
| GSM211682 |
Control R2, biological rep1 |
| GSM211683 |
Control R2, biological rep2 |
| GSM211684 |
TIGR4WT, biological rep1 |
| GSM211685 |
TIGR4WT, biological rep2 |
| GSM211686 |
TIGR4WT, biological rep3 |
| GSM211687 |
TIGR4delta-cps, biological rep1 |
| GSM211688 |
TIGR4delta-cps, biological rep2 |
| GSM211689 |
TIGR4delta-cps, biological rep3 |
| GSM211690 |
D39delta-cps_low, biological rep1 |
| GSM211691 |
D39delta-cps_low, biological rep2 |
| GSM211692 |
D39delta-cps_low, biological rep3 |
| GSM211693 |
Type2cps, biological rep1 |
| GSM211694 |
Type2cps, biological rep2 |
| GSM211695 |
Type2cps, biological rep3 |
| GSM211696 |
Control R3, biological rep1 |
| GSM211697 |
Control R3, biological rep2 |
| GSM211698 |
Control R3, biological rep3 |
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| Relations |
| BioProject |
PRJNA101655 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE8527_RAW.tar |
169.2 Mb |
(http)(custom) |
TAR (of CEL, EXP) |
| Processed data included within Sample table |
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