Goblet Cell Derived RELM-β Recruits CD4+ T Cells during Infectious Colitis to Promote Protective Intestinal Epithelial Cell Proliferation

Enterohemorrhagic Escherichia coli and related food and waterborne pathogens pose significant threats to human health. These attaching/effacing microbes infect the apical surface of intestinal epithelial cells (IEC), causing severe diarrheal disease. Colonizing the intestinal luminal surface helps segregate these microbes from most host inflammatory responses. Based on studies using Citrobacter rodentium, a related mouse pathogen, we speculate that hosts rely on immune-mediated changes in IEC, including goblet cells to defend against these pathogens. These changes include a CD4(+) T cell-dependent increase in IEC proliferation to replace infected IEC, as well as altered production of the goblet cell-derived mucin Muc2. Another goblet cell mediator, REsistin-Like Molecule (RELM)-beta is strongly induced within goblet cells during C. rodentium infection, and was detected in the stool as well as serum. Despite its dramatic induction, RELM-beta's role in host defense is unclear. Thus, wildtype and RELM-beta gene deficient mice (Retnlb(-/-)) were orally infected with C. rodentium. While their C. rodentium burdens were only modestly elevated, infected Retnlb(-/-) mice suffered increased mortality and mucosal ulceration due to deep pathogen penetration of colonic crypts. Immunostaining for Ki67 and BrDU revealed Retnlb(-/-) mice were significantly impaired in infection-induced IEC hyper-proliferation. Interestingly, exposure to RELM-beta did not directly increase IEC proliferation, rather RELM-beta acted as a CD4(+) T cell chemoattractant. Correspondingly, Retnlb(-/-) mice showed impaired CD4(+) T cell recruitment to their infected colons, along with reduced production of interleukin (IL)-22, a multifunctional cytokine that directly increased IEC proliferation. Enema delivery of RELM-beta to Retnlb(-/-) mice restored CD4(+) T cell recruitment, concurrently increasing IL-22 levels and IEC proliferation, while reducing mucosal pathology. These findings demonstrate that RELM-beta and goblet cells play an unexpected, yet critical role in recruiting CD4(+) T cells to the colon to protect against an enteric pathogen, in part via the induction of increased IEC proliferation.