Modulation of NF-kB Signaling by Immunoprobiotics

免疫益生菌对 NF-kB 信号传导的调节

• 批准号: 7674585
• 负责人: James Versalovic
• 金额: $ 45.84万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7674585
• 关键词: Affect; Bacteria; Bacterial Genes; Bioreactors; Cell Line; Cells; Coculture Techniques; Complex; Conditioned Culture Media; Down-Regulation; Ecosystem; Environment; Epithelial Cells; Gene Expression; Genes; Germ-Free; Glycerol; Glycerophospholipids; Helicobacter hepaticus; Human Microbiome; Immune; Immune response; Immune system; Immunity; Inflammation; Inflammatory; Inflammatory Response; Inflammatory disease of the intestine; Intestinal Mucosa; Intestines; Investigation; Lactobacillus; Lactobacillus reuteri; MAP Kinase Signaling Pathways; Mammalian Cell; Mammals; Metabolism; Microbe; Mitogen-Activated Protein Kinases; Modeling; Mucous Membrane; Mus; Mutagenesis; Myelogenous; Myeloid Cells; NF-kappa B; Pathway interactions; Probiotics; Production; Public Health; Regulation; Relative (related person); Research Personnel; Role; Signal Pathway; Signal Transduction; Simulate; Subgroup; Testing; base; cytokine; gut microbiota; immunoregulation; in vivo; loss of function mutation; microbial; microbial community; microbial host; microbiome; mouse model; mutant; pathogen; prebiotics; reconstitution; research study; simulation; transcription factor

DESCRIPTION (provided by applicant): The overall hypothesis of this proposal is that glycerol/glycerophospholipid metabolism is a focal point for determining the relative capacities of probiotics to suppress NF-?B and MAP kinase signaling in mammalian cells. Ultimately, suppression of key signaling pathways results in down-regulation of pro-inflammatory cytokine production, and selective depletion of activated immune cells. Targeted and random mutagenesis strategies will be used to generate insertion mutants of probiotics that gain or lose the ability to suppress NF-?B activation or MAP kinase signaling. Bioreactors with defined combinations of intestinal bacteria will be used in order to simulate gut bacterial ecosystems. Myeloid and epithelial cells will be combined in co-culture models in order to provide simplified models of the gut mucosa. Primary candidate immunoregulatory factors produced by lactobacilli include glycerophospholipid derivatives that may down-regulate pro-inflammatory responses. Finally, selected probiotic mutants will be introduced into a defined microbiota-containing mouse model in order to study effects of a defined microbiome on NF-?B activation and MAP kinase immune signaling pathways in vivo. These studies will occur within the context of well-characterized fluctuations of the intestinal microbiota so that microbial and host contributions to immunoregulation can be investigated in parallel using one mouse model. 1. Generate insertional mutants of immunoprobiotic Lactobacillus reuteri and identify key bacterial genes that regulate factors affecting NF-?B and MAP kinase signaling pathways. 2. Investigate regulation of NF-?B and MAP kinase signaling in a simulated gut mucosa by probiotics/mutants in the milieu of a defined microbiota. 3. Introduce wild type and isogenic probiotic mutants into IL-10-deficient mice with a defined microbiome in order to study probiotic effects on NF-?B and MAP kinase signaling pathways in vivo. PUBLIC HEALTH REVELANCE: The investigator seeks to understand how beneficial bacteria may regulate inflammation in the intestine. This project includes a simulation of the intestinal microbial community so that investigations take place in the context of conditions similar to the complex intestinal environment. Finally, a mouse model of intestinal inflammation will be used to explore which genes of beneficial bacteria are important for regulating intestinal immune responses in mammals.

描述（由申请人提供）：本提案的总体假设是甘油/甘油磷脂代谢是确定益生菌抑制NF-？哺乳动物细胞中的B和MAP激酶信号。最终，关键信号通路的抑制导致促炎细胞因子产生的下调，以及活化免疫细胞的选择性消耗。靶向和随机诱变策略将用于产生益生菌的插入突变体，这些突变体可以获得或失去抑制NF-？B活化或MAP激酶信号。将使用具有确定肠道细菌组合的生物反应器来模拟肠道细菌生态系统。骨髓细胞和上皮细胞将在共培养模型中结合，以提供肠道黏膜的简化模型。乳酸菌产生的主要候选免疫调节因子包括可能下调促炎反应的甘油磷脂衍生物。最后，将选定的益生菌突变体引入含有特定微生物群的小鼠模型，以研究特定微生物群对NF-？体内B活化和MAP激酶免疫信号通路。这些研究将在具有良好特征的肠道微生物群波动的背景下进行，以便微生物和宿主对免疫调节的贡献可以使用一个小鼠模型并行研究。1. 产生免疫益生菌罗伊氏乳杆菌插入突变体，鉴定调节NF-？B和MAP激酶信号通路。2. 探讨NF-?的调控作用益生菌/突变体在特定微生物群环境中模拟肠道黏膜中的B和MAP激酶信号。3. 将野生型和等基因型益生菌突变体引入具有特定微生物组的il -10缺陷小鼠，以研究益生菌对NF-？B和MAP激酶信号通路的体内研究。