Transcriptional profiling of Clostridium difficile and Caco-2 cells during infection - PubMed (original) (raw)

Transcriptional profiling of Clostridium difficile and Caco-2 cells during infection

Tavan Janvilisri et al. J Infect Dis. 2010.

Abstract

Clostridium difficile is well recognized as the most common infectious cause of nosocomial diarrhea. The incidence and severity of C. difficile infection (CDI) is increasing worldwide. Here, we evaluated simultaneously the transcriptional changes in the human colorectal epithelial Caco-2 cells and in C. difficile after infection. A total of 271 transcripts in Caco-2 cells and 207 transcripts in C. difficile were significantly differentially expressed at 1 time point during CDI. We used the gene ontology annotations and protein-protein network interactions to underline a framework of target molecules that could potentially play a key role during CDI. These genes included those associated with cellular metabolism, transcription, transport, cell communication, and signal transduction. Our data identified certain key factors that have previously been reported to be involved in CDI, as well as novel determinants that may participate in a complex mechanism underlying the host response to infection, bacterial adaptation, and pathogenesis.

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Conflict of interest statement

All authors listed in the title page do not have a commercial or other association that might pose a conflict of interest.

Figures

Figure 1

Figure 1

Infection of Caco-2 cells with C. difficile. (A) Quantification of Caco-2 cell viability at different times post-infection. All assays were conducted in triplicate and repeated independently three times. Cell viability is as expressed as the percentage of survival of the control wells. Results are expressed as means ± 1 standard deviation for the replicate experiments, and the Student t test was used for statistical analysis of the data. Significant difference from the control (p < 0.05) is indicated by an asterisk. Caco-2 cells under anaerobic conditions at different time points without the infection were also evaluated but they were not significantly different from the control cells. (B) Photomicrographs of infected Caco-2 cells with C. difficile at 30, 60 and 120 min post-infection.

Figure 2

Figure 2

Transcriptional dialogue between Caco-2 cells and C. difficile during infection. (A) The number of up- or down-regulated genes after 30, 60, and 120 min p.i., as compared to the expression levels at the time of infection. (B, C) Hierarchical clustering analysis of differentially expressed genes in Caco-2 cells and C. difficile during infection. Genes identified to be significantly differentially expressed at 30, 60 or 120 min in Caco-2 or C. difficile cells p.i. relative to in vitro growth. Genes significantly different with _p_-value < 0.05 after the infection were pooled and used to create heatmaps for (B) Caco-2 cells, and (C) C. difficile. Genes are ordered in rows, conditions as columns. Red color indicates genes induced post-infection vs. prior to infection (fold change); green color denotes repression.

Figure 3

Figure 3

Functional annotation of genes in Caco-2 cells and C. difficile, which are differentially expressed between infected and uninfected conditions. All differentially expressed genes were annotated using generic GO-slim for biological process.

Figure 4

Figure 4

Validation of microarray data by qRT-PCR. Gene expression changes in infected versus uninfected cells measured by microarray analysis or qRT-PCR are compared. Data are plotted as log2 ratios of microarray data (x-axis) compared to those of qRT-PCR (y-axis).

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