IL-27 induces LL-37/CRAMP expression from intestinal epithelial cells: implications for immunotherapy of Clostridioides difficile infection

AbstractClostridioides difficile spores produced during infection are important for the recurrence of the disease. Here, we show that C. difficile spores gain entry into the intestinal mucosa via pathways dependent on host fibronectin-α5β1 and vitronectin-αvβ1. The exosporium protein BclA3, on the spore surface, is required for both entry pathways. Deletion of the bclA3 gene in C. difficile, or pharmacological inhibition of endocytosis using nystatin, leads to reduced entry into the intestinal mucosa and reduced recurrence of the disease in a mouse model. Our findings indicate that C. difficile spore entry into the intestinal barrier can contribute to spore persistence and infection recurrence, and suggest potential avenues for new therapies.

Download Full-text

E-cadherin redistribution mediated by Clostridioides difficile toxins increases spore-association to adherens junctions

10.1101/2021.09.01.458577 ◽

2021 ◽

Author(s):

Pablo Castro-Cordova ◽

Macarena Otto-Medina ◽

Borden Lacy ◽

Daniel Paredes-Sabja

Keyword(s):

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Adherens Junctions ◽

Immunogold Labelling ◽

Intestinal Epithelial ◽

Clostridioides Difficile ◽

Spore Adherence ◽

E Cadherin

Nearly ~20% of patients with C. difficile infection (CDI) manifest recurrence of CDI (R-CDI). During CDI, C. difficile forms spores essential for R-CDI. Interactions of C. difficile spores with intestinal epithelial cells (IECs) contribute to R-CDI. However, this interaction remains poorly understood. Here, we provide evidence that C. difficile spores interact with E-cadherin, contributing to spore-adherence and internalization into IECs. C. difficile toxins TcdA/TcdB lead to adherens junctions opening and increase spore-adherence to IECs. Confocal micrographs demonstrate that C. difficile spores associate with accessible E-cadherin; spore-E-cadherin association increases upon TcdA/TcdB intoxication. The presence of anti-E-cadherin antibodies decreased spore adherence and entry into IECs. By ELISA, immunofluorescence, and immunogold labelling, we observed that E-cadherin binds to C. difficile spores, specifically to the hair-like projections of the spore. Overall, these results expand our knowledge of how C. difficile spores bind to IECs and how toxin-mediated damage affects spore interactions with IECs.

Download Full-text

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

10.1101/2020.09.11.291104 ◽

2020 ◽

Author(s):

Pablo Castro-Córdova ◽

Paola Mora-Uribe ◽

Rodrigo Reyes-Ramírez ◽

Glenda Cofré-Araneda ◽

Josué Orozco-Aguilar ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Rational Design ◽

Intestinal Epithelial Cells ◽

Intestinal Barrier ◽

Recurrent Infection ◽

Intestinal Epithelial ◽

Clostridioides Difficile ◽

Infection Recurrence

AbstractClostridioides 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-α5β1 and vitronectin-αvβ1 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.

Download Full-text

Three-dimensional organization and functional relationships of endocytotic compartments in pig intestinal epithelial cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123283 ◽

1992 ◽

Vol 50 (1) ◽

pp. 576-577

Author(s):

Julian P. Heath ◽

Buford L. Nichols ◽

László G. Kömüves

Keyword(s):

Electron Microscopy ◽

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Three Dimensional ◽

Intestinal Epithelial ◽

Cell Surfaces ◽

Basal Surface ◽

Functional Relationships

The newborn pig intestine is adapted for the rapid and efficient absorption of nutrients from colostrum. In enterocytes, colostral proteins are taken up into an apical endocytotic complex of channels that transports them to target organelles or to the basal surface for release into the circulation. The apical endocytotic complex of tubules and vesicles clearly is a major intersection in the routes taken by vesicles trafficking to and from the Golgi, lysosomes, and the apical and basolateral cell surfaces.Jejunal tissues were taken from piglets suckled for up to 6 hours and prepared for electron microscopy and immunocytochemistry as previously described.

Download Full-text