Host Response

主持人回应

基本信息

批准号：
8026693
负责人：
Alessio Fasano
金额：
$ 31.73万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-20 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8026693
关键词：
5 year old Affect Age Antibodies Antigens Area Bacteria Biochemistry Biological Markers Biology Child Clinical Complement Complex Country Diarrhea Disease Electrolyte Balance Enteral Environment Epithelial Equilibrium Escherichia coli Infections Feces Gastrointestinal Physiology Gastrointestinal tract structure Genetic Genomics Homeostasis Immune response Immune system Immunoglobulin A Immunoglobulin G Immunologic Markers Immunology Infection Inflammation Mediators Inflammatory Intestinal Motility Intestinal Secretions Intestines Ions Knockout Mice Knowledge Lead Liquid substance Measures Mediating Microbe Microbiology Molecular Movement Mucosal Immune Responses Mucous Membrane Mus Mutant Strains Mice Nutrient Outcome Pathogenesis Pathway interactions Pattern recognition receptor Permeability Physiological Polymorphism Analysis Process Production Proteinase-Activated Receptors Research Role Sampling Shigella Shigella flexneri Signal Transduction Single Nucleotide Polymorphism Specimen TLR4 gene TLR5 gene Testing Tight Junctions Tissues Virulence Factors Wild Type Mouse absorption adaptive immunity cell motility cell type cytokine enteric pathogen enteropathogenic Escherichia coli gastrointestinal human tissue immune function intestinal epithelium killings laser capture microdissection mast cell mutant pathogen response trafficking vaccine development

项目摘要

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.

腹泻疾病是5岁以下儿童的主要杀手。这些感染感染代表了微生物与宿主之间的平衡。宿主对肠感染的反应包括在肠上皮和粘膜/全身免疫系统中看到的反应。该项目将使用小鼠肠道组织和人类临床标本检查两种类型的反应。具体的目标1将表征安装在Ussing Chamber和Microsnapwells中的小鼠上皮组织对志贺氏菌感染和肠病大肠杆菌（EPEC）的感染，特别着重于肠壁屏障功能，短路电流和上皮细胞因子的产生。将测试一些在特定毒力因子中有缺陷的同源突变体，以确定这些因素对所见反应的贡献。与将采用野生型小鼠和各种细菌突变体的AIM 1相比，特定的AIM 2将采用野生型细菌和各种突变小鼠菌株来确定特定模式识别受体（PRR）在肠粘膜对志贺氏菌和EPEC的反应中的作用。肠道组织将从野生型中分离出来，并淘汰缺乏MyD88，TLR4，TLR5，Nodi，Nodi，Nod2，Par2或肥大细胞的小鼠，并在Shigella and Epec中暴露于体内。在不同小鼠系中的肠粘膜功能和免疫反应的比较将表征这些PRR在EPEC和Shigella引起的感染中的作用。特定的目标3将检查与经常国家儿童中与志贺氏菌和EPEC感染有关的GL道中的适应性免疫和免疫学标记。将分析来自记录的志贺氏菌和EPEC感染儿童的粪便标本，以分析病原体特异性自适应反应，包括SIGA，IGA和IgG，以及特异于志贺氏菌和EPEC抗原的IgG。还将分析粪便细胞因子和炎症分子，并寻求与临床结局相关的相关性。