Abstract
The aim of this study was to test the hypothesis that Bacillus subtilis antagonises enterotoxic Escherichia coli (ETEC) infection through mitogen-activated protein kinases (MAPK) signaling pathways. In vitro studies were performed in which ETEC-infected Caco-2 cultured human intestinal cells were first incubated with B. subtilis and then ETEC adhesion and MAPK activation were determined. Incubation with B. subtilis was found to reduce ETEC adhesion in Caco-2 cells by 58–72 % in the adhesive experiments (competition, exclusion, and displacement assays). ETEC was able to induce extracellular signal-regulated kinases 1 and 2 (ERK1/2) and p38 MAPK activation, but not c-Jun amino-terminal kinase (JNK) activation, in Caco-2 cells. ETEC-induced phosphorylation of ERK1/2, but not of p38, was inhibited significantly in ETEC-infected Caco-2 cells treated with B. subtilis. These findings suggest that B. subtilis is able to inhibit ETEC infection through blocking ETEC-induced ERK1/2 activation in Caco-2 cells. The data could provide a rationale for the clinical application of B. subtilis in enteric pathogenic infection.
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This work was supported by grants no. 30871858 from the National Science Grant of China and no. BE200830155 from the support programme of Jiangsu province.
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Supplementary Fig. S1
Activation of extracellular signal-regulated kinases (ERK1/2) by ETEC is regulated at different time points. Caco-2 cells were infected with ETEC for the times indicated. Cell lysates were prepared at each time point and analysed by Western blotting for the presence of phosphorylated ERK (p-ERK). The blots were then stripped and probed again with antibodies for total ERK. The images were analysed using Quantity One software, and the mean density of each band was measured. The density of the phosphorylated ERK band was divided by the density of the corresponding total ERK band to obtain the normalised band density, which is indicated below each lane. The results shown are representative of three separate experiments (the band densities presented refer specifically to the Western blots shown)
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Ye, X., Li, P., Yu, Q. et al. Bacillus subtilis inhibition of enterotoxic Escherichia coli-induced activation of MAPK signaling pathways in Caco-2 cells. Ann Microbiol 63, 577–581 (2013). https://doi.org/10.1007/s13213-012-0506-8
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DOI: https://doi.org/10.1007/s13213-012-0506-8