Mendoza-Leon, M. J. | bioRxiv Science

bioRxiv2021

A ligated intestinal loop mouse model protocol to study the interactions of Clostridioides difficile spores with the intestinal mucosa during aging.

The interaction of the Clostridioides difficile spores with the intestinal mucosa contribute to the persistence and recurrence of the infection. Advanced age is one of the main risk factors to manifest C. difficile infection and recurrence. However, the interaction of C. difficile spores with the intestinal mucosa during aging has not been evaluated. In the present work, we provide a detailed protocol with all the critical information to perform an intestinal ligated loop. Using this technique in a mouse model, we evaluated C. difficile spore adherence and internalization to the ileum and colonic mucosa during aging. Consequently, our data suggest that spore internalization in the ileum and colonic mucosa is higher in elderly than in adults or young mice. Also, our data suggest that spore-adherence to the ileum and colonic mucosa decreases with aging.

microbiology↗

bioRxiv2020

Clostridioides difficile spore-entry into intestinal epithelial cells contributes to recurrence of the disease

Clostridioides difficile spores produced during infection are essential for the recurrence of the disease. However, how C. difficile spores persist in the intestinal mucosa to cause recurrent infection remains unknown. Here, we show that C. difficile spores gain entry into the intestinal mucosa via fibronectin-5{beta}1 and vitronectin-v{beta}1 specific-pathways. The spore-surface exosporium BclA3 protein is essential for both spore-entry pathways into intestinal epithelial cells. Furthermore, C. difficile spores of a bclA3 isogenic mutant exhibited reduced entry into the intestinal mucosa and reduced recurrence of the disease in a mouse model of the disease. Inhibition of C. difficile spore-entry led to reduced spore-entry into the intestinal epithelial barrier and recurrence of C. difficile infection in vivo. These findings suggest that C. difficile spore-entry into the intestinal barrier is a novel mechanism of spore-persistence that can contribute to infection recurrence and have implications for the rational design of therapies.

microbiology↗