Host Response

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Diarrheal diseases are a major killer of children under the age of 5. These infections infections represent a balance between the microbe and the host. Host responses to enteric infections include those seen in the intestinal epithelium and in the mucosal/systemic immune system. This project will examine both types of responses using mouse intestinal tissue and human clinical specimens. Specific Aim 1 will characterize the response of mouse epithelial tissue mounted in Ussing chambers and microsnapwells to infection with Shigella and enteropathogenic E. coli (EPEC) with particular emphasis on intestinal barrier function, short circuit current, and epithelial cytokine production. Several isogenic mutants defective in specific virulence factors will be tested to determine the contribution of these factors to the responses seen. In contrast to Aim 1, which will employ wild type mice and various bacterial mutants, Specific Aim 2 will employ wild type bacteria and various mutant mouse strains to establish the role of specific pattern recognition receptors (PRRs) in the intestinal mucosal response to Shigella and EPEC. Intestinal tissue will be isolated from wild type and knock out mice deficient in MyD88, TLR4, TLR5, Nodi, Nod2, PAR2 , or mast cells and exposed ex vivo to Shigella and EPEC. Comparison of the intestinal mucosal function and immune responses in the different mouse lines will characterize the role of these PRRs in infections due to EPEC and Shigella. Specific Aim 3 will examine adaptive immunity and immunological markers in the Gl tract that are associated with Shigella and EPEC infection in children from endemic countries. Stool specimens from children with documented Shigella and EPEC infections in the GEMS study will be analyzed for pathogen specific adaptive responses including sIgA, IgA, and IgG specific for Shigella and EPEC antigens. Fecal cytokines and inflammatory molecules will also be analyzed and correlations sought with clinical outcomes.