P063 Host-mycobiome interactions during the resolution of inflammation in experimental colitis

De Salvo, C.(1)*;Wargo, H.(1);Vidmar, K.(1);Di Martino, L.(2);Mahabaleshwar, G.(1);Cominelli, F.(2);Ghannoum, M.(2);Pizarro, T.(1);

(1)Case Western Reserve University- School of Medicine, Pathology, Cleveland, United States;(2)Case Western Reserve University- School of Medicine, Digestive Health Research Institute, Cleveland, United States;


Recent studies have documented the complexity of the intestinal fungal community (‘mycobiome’) in mice, and clinical and experimental observations have shown that the mycobiome influences both gut health and disease, e.g., IBD. In fact, prior studies have shown that CD patients, compared to healthy controls, harbor higher levels of intestinal Candida tropicalis (Ct), which is the major fungal species detected in the colons of colitic mice after dextran sodium sulfate (DSS) challenge. Moreover, proteins encoded by genes within IBD susceptibility loci, such as the pattern recognition receptor (PRR), NOD2, are known not only to recognize bacterial components, but also a fungal cell wall element, chitin. In fact, the role of PRRs in regulating immunity against intestinal fungi, and how fungi influence IBD remains poorly defined.


We challenged DSS colitic wildtype (WT) and Nod2-/- mice with Ct 2 days before DSS administration and subsequently on day 0, 3 and 6. Bone marrow-derived macrophages (BMDMs) from untreated mice were administered chitin in vitro.


Our data confirms previous studies that Ct challenge does not exacerbate colitis in DSS-treated C57BL/6 WT mice, however, Ct-infected Nod2-/- mice possess a higher fungal burden and exhibit worse colitis symptoms, such as weight loss, decreased stool consistency, and presence of blood in  stools, vs. Ct-infected WT mice, indicating an essential and protective role for NOD2 during colitis recovery after Ct challenge. Our results also show that Ct-infected Nod2-/- mice display a marked reduction in colonic Il22 and Il17, which are cytokines previously reported to be important in maintaining epithelial barrier integrity during DSS colitis. These data were confirmed in colons of Ct infected DSS challenged, ileitis prone SAMP1/YitFc (SAMP) mice NOD2 deficient. Moreover, our in vitro data show, a decrease in Il1ß and Il23 in BMDM from SAMP Nod2-/- mice compared to WT after 2h of exposure to chitin. IL-17+ innate lymphoid cells (ILCs) are also known to control fungal burden during opportunistic fungal infections, and interestingly, our findings indicate that Ct infection of Nod2-/- vs. WT mice results in a decreased frequency of mesenteric lymph node–derived type 3 ILCs (ILC3s), suggesting that delay in fungal clearance and recovery in Nod2-/-mice may be due to the inability to mount protective type 3 immune responses.


Taken together, the data collected suggests that NOD2 is essential to maintain gut mycobiome homeostasis and drives protective innate immune responses, via a macrophage-mediated ILC3 recruitment and IL-17 mechanism, by preventing the overgrowth of opportunistic fungi that may contribute to chronic intestinal inflammation, such as that observed in CD.