Human intestinal enteroids as a model of Clostridioides difficile-induced enteritis.

Human intestinal enteroids as a model of Clostridioides difficile-induced enteritis. Am J Physiol Gastrointest Liver Physiol. 2020 Mar 30;: Authors: Engevik MA, Danhof HA, Chang-Graham AL, Spinler JK, Engevik KA, Hermann B, Endres BT, Garey KW, Hyser JM, Britton RA, Versalovic J Abstract Clostridioides difficile is an important nosocomial pathogen that produces toxins to cause life-threatening diarrhea and colitis. Toxins bind to epithelial receptors and promote the collapse of the actin cytoskeleton. C. difficile toxin activity is commonly studied in cancer-derived and immortalized cell lines. However, the biological relevance of these models are limited. Moreover, no model is available for examining C. difficile-induced enteritis, an understudied health problem. We hypothesized human intestinal enteroids (HIEs) express toxin receptors and provide a new model to dissect C. difficile cytotoxicity in the small intestine. We generated biopsy-derived jejunal HIE and Vero cells which stably express LifeAct-Ruby, a fluorescent label of F-actin, to monitor actin cytoskeleton rearrangement by live-cell microscopy. Imaging analysis revealed that toxins from pathogenic C. difficile strains elicited cell rounding in a strain-dependent manner and HIEs were tenfold more sensitive to toxin A (TcdA) than toxin B (TcdB). By qPCR we paradoxically found that HIEs expressed greater quantities of toxin receptor mRNA and yet exhibited decreased sensitiv...
Source: American Journal of Physiology. Gastrointestinal and Liver Physiology - Category: Physiology Authors: Tags: Am J Physiol Gastrointest Liver Physiol Source Type: research