Use of an in-vitro polarized cell culture model to study the translocation of Clostridium difficile toxins
Clostridium difficile is the etiologic agent of antibiotic-associated diarrhea; the most common form of nosocomial infectious diarrhea. C. difficile produces two large molecular weight protein exotoxins; toxins A and B. In this study, a polarized tissue culture model employing Caco-2 cells grown on...
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| Format: | Others |
| Language: | en en_US |
| Published: |
2007
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| Online Access: | http://hdl.handle.net/1993/1997 |
| Summary: | Clostridium difficile is the etiologic agent of antibiotic-associated diarrhea; the most common form of nosocomial infectious diarrhea. C. difficile produces two large molecular weight protein exotoxins; toxins A and B. In this study, a polarized tissue culture model employing Caco-2 cells grown on Transwell inserts was established to study the translocation of purified Clostridium difficile toxins A and B. Clostridium difficile toxins were $\sp{125}$I labeled and inoculated onto confluent polarized Caco-2 cell monolayers to study translocation dynamics. The ability of toxins A and B to enhance translocation of lipopolysaccharide was also assessed. Electrical resistance measurements were utilized to monitor monolayer confluence and tight junction integrity. Sera from patients afflicted with C. difficile-associated diarrhea were analyzed for circulating cytotoxin, as well as, serum neutralizing antibodies. Synsorb CD and Cholestyramine were also examined for their ability to inhibit cytotoxic effects on Caco-2 cells and Human foreskin fibroblasts. (Abstract shortened by UMI.) |
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