Yersinia Pseudotuberculosis YopE Prevents Uptake by M Cells and Instigates M Cell Extrusion in Human Ileal Enteroid-derived Monolayers

Many pathogens use M cells to access the underlying Peyer's patches and spread to systemic sites via the lymph as demonstrated by ligated loop murine intestinal models. However, the study of interactions between M cells and microbial pathogens has stalled due to the lack of cell culture systems. To overcome this obstacle, we use human ileal enteroid-derived monolayers containing five intestinal cell types including M cells to study the interactions between the enteric pathogen, (), and M cells. The type three secretion system (T3SS) effector Yops inhibit host defenses including phagocytosis and are critical for colonization of the intestine and Peyer's patches. Therefore, it is not understood how traverses through M cells to breach the epithelium. By growing under two physiological conditions that mimic the early infectious stage (low T3SS-expression) or host-adapted stage (high T3SS-expression), we found that large numbers of specifically associated with M cells, recapitulating murine studies. Transcytosis through M cells was significantly higher by expressing low levels of T3SS, because YopE and YopH prevented uptake. YopE also caused M cells to extrude from the epithelium without inducing cell-death or disrupting monolayer integrity. Sequential infection with early infectious stage reduced host-adapted association with M cells. These data underscore the strength of enteroids as a model by discovering that Yops impede M cell function, indicating that early infectious stage more effectively penetrates M cells while the host may defend against M cell penetration of host-adapted .