Furthermore, we did not find any genes with similar sequence to the CDTB gene using a BLAST search of the published C. concisus genome (NCBI accession number NC_009802), indicating that other factors (i.e. selleck inhibitor opposed to the CDT) may be responsible. The role that Campylobacter-induced epithelial cell death plays in pathogenesis is currently poorly understood;
hence, the clinical significance of these findings for C. concisus remains to be determined. Metabolic activity can be measured using the MTT assay in which metabolically active epithelial cells reduce a yellow tetrazolium salt (MTT) to purple formazan crystals that can be spectrophotometrically quantified. All of the isolates that we examined, except one https://www.selleckchem.com/products/AC-220.html isolate that caused epithelial sloughing (CHRB6), induced higher MTT values (> 130%) than the control, indicating that epithelial metabolic activity is increased by C. concisus. Some clinical strains of C. jejuni have also been reported to cause similar increases in epithelial MTT values [31]. Given the short incubation period for the MTT assay, we conclude that the increased values most likely reflect selleckchem an increase in metabolic activity due to cellular stress rather than an increase in epithelial cell numbers due to proliferation. The observed
correlation between metabolic activity and DNA fragmentation may be a consequence of the increased energy demands required to sustain the apoptotic process (i.e., apoptotic DNA fragmentation is an ATP-dependent process [32]). The chemokine, IL-8 is a major mediator of inflammation. In the current study, all C. concisus isolates induced transcription of IL-8 in epithelial monolayers (> 2-fold) as has been previously reported for C. jejuni [19] and C. concisus [33]. Campylobacter jejuni induces
epithelial IL-8 secretion by at least two independent mechanisms, one of which requires invasion and the other that is CDT-dependent [19, 34]. We observed see more that induction of IL-8 transcription by C. concisus was not correlated with invasion. Man et al. also recently showed that three C. concisus strains stimulated production of IL-8 in intestinal epithelial irrespective of their invasive ability [33]. Thus in contrast to C. jejuni, it appears that factors other than invasion or CDT (which appears to be lacking in this species) are responsible for the up-regulation of IL-8 incited by C. concisus. The observation that expression of IL-8 mRNA was greater in epithelial cells treated with isolates from AFLP cluster 1 compared to isolates from cluster 2 was unexpected and suggests that these isolates may have pathogenic potential. We identified genes encoding S-layer RTX and the zonnula occludins toxin in some of the isolates, confirming initial reports of these toxin genes in C. concisus [21]. Surprisingly, the zot gene was more prevalent in isolates from healthy (80%) compared to diarrheic (22%) humans.