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Vet Pathol. 2011 Mar;48(2):369-80. doi: 10.1177/0300985810380395. Epub 2010 Sep 22.

The attachment, internalization, and time-dependent, intracellular distribution of Clostridium difficile toxin A in porcine intestinal explants.

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Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011, USA.


Toxin A (TcdA), secreted by toxigenic strains of Clostridium difficile, produces lesions typical of C. difficile-associated disease (CDAD) in susceptible mammal species. Porcine colon explants maintained for 2 hours with TcdA developed severe lesions characterized by cell swelling, swelling of mitochondria and other organelles, distension of cytoplasmic vesicles, expansion of paracellular spaces, apoptosis, and necrosis. Severity of lesions was proportional to the dosage of toxin. No lesions were present in uninoculated control tissues after 2 hours. Receptor-mediated endocytosis is the keystone event in the pathogenesis of the toxin, and susceptibility of a given species is thought to depend on the presence of receptors in intestinal epithelial cells. The fate of TcdA applied to viable colon explants was determined by transmission electron microscopy in an anti-toxin-labeled gold assay. At 5 minutes postinoculation, the presence of TcdA was indicated at the membrane of microvilli or in the cytoplasm of epithelial cells. TcdA was also indirectly observed within endosomes or attached at their margin. A 30-minute inoculation period was associated with many more gold particles labeling structures inside the cell, although some were still attached to microvilli. Within the cell, most TcdA was associated with mitochondria of epithelial cells, but some gold particles decorated the nuclei. Endothelial cells of the lamina propria had evidence of TcdA at both their lumenal and basal aspects, as well as in the cytoplasm and, occasionally, nuclei. Gold particles also labeled the lumen of such vessels as well as leucocytes in blood vessels and the lamina propria.

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