Translocation of Clostridium difficile Toxin B across Polarized Caco-2 Cell Monolayers Is Enhanced by Toxin A

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dc.contributor.authorDu, Tim
dc.contributor.authorAlfa, Michelle J
dc.date.accessioned2016-06-08T19:07:28Z
dc.date.available2016-06-08T19:07:28Z
dc.date.issued2004-1-1
dc.date.updated2016-06-07T07:00:09Z
dc.description.abstractClostridium difficile is the etiological agent of antibiotic-associated diarrhea; the most common form of nosocomial infectious diarrhea. The basis for the shock-like systemic symptoms observed in severe cases of this infection are not known. It is hypothesized that the invasion of C difficile toxins A and/or B from the gut mucosa may contribute to these symptoms.A polarized tissue culture model employing Caco-2 cells grown on transwell inserts was established to study the translocation of purified C difficile toxins A and B. C difficile toxins were 125I labelled and inoculated onto confluent polarized Caco-2 cell monolayers to study translocation dynamics. Electrical resistance measurements were used to monitor monolayer confluence and tight junction integrity. Samples were taken from the apical and basal sides of the insert, as well as the insert itself, and tested using the human foreskin fibroblasts cell cytotoxicity assay to monitor partitioning of the radiolabelled toxins.Toxin A produced a 50% reduction in electrical resistance in 3 h whereas the same concentration of toxin B required at least 7 h to achieve the same effect. Both toxins A and B were able to translocate across confluent monolayers of Caco-2 cells. The combination of toxin A and B together was synergistic with respect to promoting the translocation of toxin B. Although the addition of toxin A resulted in a 100% increase in the amount of toxin B able to translocate, no increases in toxin A translocation were observed. These findings suggest a model of pathogenesis in which C difficile toxin A facilitates the translocation of toxin B from the gut into submucosal areas where it may play a role in inflammatory damage.
dc.description.versionPeer Reviewed
dc.identifier.citationTim Du and Michelle J Alfa, “Translocation of Clostridium difficile Toxin B across Polarized Caco-2 Cell Monolayers Is Enhanced by Toxin A,” Canadian Journal of Infectious Diseases, vol. 15, no. 2, pp. 83-88, 2004. doi:10.1155/2004/292580
dc.identifier.urihttp://dx.doi.org/10.1155/2004/292580
dc.identifier.urihttp://hdl.handle.net/1993/31423
dc.language.rfc3066en
dc.rightsopen accessen_US
dc.rights.holderCopyright © 2004 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
dc.titleTranslocation of Clostridium difficile Toxin B across Polarized Caco-2 Cell Monolayers Is Enhanced by Toxin A
dc.typeJournal Article
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