Establishing an Enteric Infection: Complex and highly regulated interactions are required to keep the peace between the bacteria that reside in our gut and the immune system. How do pathogenic bacteria, such as the strains of Escherichia coli that cause gastroenteritis, get a foothold to establish an infection, and what is the role of resident bacteria in this process? Kamada et al. (p. 1325 , published online 10 May; see the Perspective by Sperandio ) infected mice orally with Citrobacter rodentium and found that mice with normal commensal microflora, which were better able to contain the infection than mice that lacked the commensals, which were not able to clear the infection.

Despite recent advances in our understanding of the pathogenesis of attaching and effacing (A/E) Escherichia coli infections, the mechanisms by which the host defends against these microbes are unclear. The goal of this study was to determine the role of goblet cell-derived Muc2, the major intestinal secretory mucin and primary component of the mucus layer, in host protection against A/E pathogens. To assess the role of Muc2 during A/E bacterial infections, we inoculated Muc2 deficient (Muc2^−/−) mice with Citrobacter rodentium, a murine A/E pathogen related to diarrheagenic A/E E. coli. Unlike wildtype (WT) mice, infected Muc2^−/− mice exhibited rapid weight loss and suffered up to 90% mortality. Stool plating demonstrated 10–100 fold greater C. rodentium burdens in Muc2^−/− vs. WT mice, most of which were found to be loosely adherent to the colonic mucosa. Histology of Muc2^−/− mice revealed ulceration in the colon amid focal bacterial microcolonies. Metabolic labeling of secreted mucins in the large intestine demonstrated that mucin secretion was markedly increased in WT mice during infection compared to uninfected controls, suggesting that the host uses increased mucin release to flush pathogens from the mucosal surface. Muc2 also impacted host-commensal interactions during infection, as FISH analysis revealed C. rodentium microcolonies contained numerous commensal microbes, which was not observed in WT mice. Orally administered FITC-Dextran and FISH staining showed significantly worsened intestinal barrier disruption in Muc2^−/− vs. WT mice, with overt pathogen and commensal translocation into the Muc2^−/− colonic mucosa. Interestingly, commensal depletion enhanced C. rodentium colonization of Muc2^−/− mice, although colonic pathology was not significantly altered. In conclusion, Muc2 production is critical for host protection during A/E bacterial infections, by limiting overall pathogen and commensal numbers associated with the colonic mucosal surface. Such actions limit tissue damage and translocation of pathogenic and commensal bacteria across the epithelium. Enteropathogenic E. coli (EPEC) and Enterohemorrhagic E. coli (EHEC) are important causes of diarrheal disease and other serious complications worldwide. Despite many studies addressing the pathogenic strategies used by these microbes, how the host protects itself from these pathogens is poorly understood. A critical question we address here is whether the thick mucus layer that overlies the intestinal surface plays a role in host protection. Since EPEC and EHEC do not infect mice efficiently, we used a related mouse pathogen called Citrobacter rodentium to infect and compare responses between wildtype mice and Muc2-deficient mice, which are defective in mucus production. We show that Muc2-deficient mice are extremely susceptible to C. rodentium infection-induced mortality and disease. Muc2-deficient mice were also colonized faster and had higher pathogen burdens throughout the experiment. Resident (non-pathogenic) bacteria were found to interact with C. rodentium and host tissues in Muc2-deficient mice, indicating Muc2 regulates all forms of intestinal microbiota at the gut surface. Deficiency in mucus production also contributed to increased leakiness of the gut, which allowed microbes to enter mucosal tissues. Our study shows that Muc2-dependent mucus production is critical for effective management of both pathogenic and non-pathogenic bacteria during infection by an EPEC/EHEC-like pathogen.

Bacterial pathogenicity islands (PAI) often encode both effector molecules responsible for disease and secretion systems that deliver these effectors to host cells. Human enterohemorrhagic Escherichia coli (EHEC), enteropathogenic E. coli , and the mouse pathogen Citrobacter rodentium (CR) possess the locus of enterocyte effacement (LEE) PAI. We systematically mutagenized all 41 CR LEE genes and functionally characterized these mutants in vitro and in a murine infection model. We identified 33 virulence factors, including two virulence regulators and a hierarchical switch for type III secretion. In addition, 7 potential type III effectors encoded outside the LEE were identified by using a proteomics approach. These non-LEE effectors are encoded by three uncharacterized PAIs in EHEC O157, suggesting that these PAIs act cooperatively with the LEE in pathogenesis. Our findings provide significant insights into bacterial virulence mechanisms and disease.