Stroke-induced Gut Microbiota Dysbiosis Regulates Microfold Cells In Peyer's Patches

Microfold or membranous cells (M cells) are specialized antigen sampling cells residing in the epithelium of Peyer’s patches (PPs), the gut-associated lymphoid tissue in the small intestine. M cells are in continuous crosstalk with luminal microbes and host immune cells. The detrimental shift of the microbiota seen with aging and after stroke contribute to bacterial antigen translocation. This axis has emerged as an epicenter for post-stroke immune dysfunction and systemic infection. The role of M cells in the PPs as an initiation site for host mucosal immunity after stroke is undefined. Hypothesis: Stroke-induced gut dysbiosis and M cell ablation leads to impaired antigen sampling mechanisms and clearance of translocating bacteria in PPs after stroke. We used a 60-minute reversible middle cerebral artery occlusion model in young (8-10 wks) C57BL/6 male mice to investigate how brain ischemia affects M cells in the PPs. We performed microbiota transplants from the cecal contents of stroke mice to naïve age-matched recipients via oral gavage for three consecutive days before tissue harvest on day four. We determined that stroke-induced changes in gut microbiota alone can cause M cell dysfunction. We found that both the number of PPs and M cells decrease 24 hours after stroke (n=8/gp, p=0.0104 and p=0.0054, respectively). Our imaging studies revealed disruption of tissue architecture and reduction in size of PPs after stroke. Microbiota transplant from stroke mice cecum to naïve recipients showed a similar effect on the number of PPs and M cells (n=10/gp, p=0.0568 and p=0.0299 ). The decrease in the number of M cells after microbiota transplantation was associated with immune dysregulation in the PPs, such as a reduction in the number of regulatory T cells (n=5/gp, p=0.0084 ). This is the first study that specifically examined M cells in a mouse model of stroke. Our results show that 1) stroke reduces the number of PPs and M cells and 2) stroke-induced gut dysbiosis can independently reduce the number of PPs and likely M cells and may regulate gut-originated immune responses after stroke. Future studies are needed to understand the effects of stroke-induced dysbiosis on M cell-mediated antigen processing in the gut and their immunoregulatory functions.