Ubiquitylation and the Regulation of Immune Homeostasis

泛素化与免疫稳态的调节

基本信息

批准号：
8536787
负责人：
Robert Cohen
金额：
$ 46.74万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8536787
关键词：
A20 protein Alleles Anti-Inflammatory Agents Anti-inflammatory Antigens Binding Biochemical Biochemistry C-terminal CD11 Antigens Cell physiology Cells Cellular biology Cleaved cell Cysteine Dendritic Cells Disease Enzymes Exhibits Gene Targeting Genes Goals Health Homeostasis ITGAX gene Immune Immune response In Vitro Inflammatory Knock-in Mouse Laboratories Ligands Link Mediating Mouse Strains Mus Mutation N-terminal Natural Immunity Physiological Play Polyubiquitin Proteins Public Health Reagent Receptor Signaling Recruitment Activity Regulation Role Signal Pathway Signal Transduction Signaling Protein Specificity TNF gene TRAF6 gene Toll-like receptors Transgenic Mice Ubiquitin Ubiquitination Work Zinc Fingers in vivo novel peripheral tolerance protein protein interaction response therapy development ubiquitin isopeptidase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Dendritic cells (DCs) are innate immune cells that regulate both immune homeostasis as well as antigen driven immune responses. The functions of DCs are highly regulated by signals initiated by Toll-like receptors (TLRs). Consequently, intracellular molecules that regulate TLR signaling pathways are pivotal for regulating host immune responses. We have recently found that A20 is a ubiquitin modifying enzyme that restricts toll-like receptor (TLR) induced signals. We are studying the roles of A20 expression in DCs in regulating immune homeostasis and responses. To accomplish this, we have generated two strains of gene targeted mice: one that disrupts the A20 gene in all cells and one in which a "floxed" allele of A20 has been targeted to the endogenous A20 locus. We have interbred the "floxed" mice with a novel strain of CD11-Cre transgenic mouse line that expresses Cre enzyme with high selectivity and specificity in DCs. These compound A20flox/flox CD11c-Cre mice will thus provide us with a very powerful and novel reagent for studying the role of A20 expression in DCs in regulating immune homeostasis and immune responses. We propose to use these A20flox/flox CD11-cre compound mice, as well as cells derived from both strains of mice, to determine how A20 expression in DCs regulates their responses to TLR ligands in vitro and in vivo. The first aim will focus upon understanding the cell biology and biochemistry underlying A20's roles in regulating TLR signaling in DCs, and will elucidate the cell-autonomous functions of DCs that are regulated by A20. The second aim will utilize A20flox/flox CD11c-Cre mice to determine how A20 expression in DCs regulates their homeostasis and activation in vivo. This second aim will also interrogate how A20 expression in DCs regulates peripheral tolerance and immune responses. PUBLIC HEALTH RELEVANCE: This project focuses on a potent endogenous anti-inflammatory protein called A20, and how it regulates dendritic cell functions in vivo. As dendritic cells are powerful regulators of immune responses, and as A20 plays critical roles in regulating dendritic cell functions, understanding how A20 regulates dendritic cells and immune responses could significantly enhance the development of therapies for inflammatory diseases and thus have major benefits for public health.

描述（由申请人提供）：树突状细胞（DC）是调节免疫稳态以及抗原驱动的免疫反应的先天免疫细胞。 DC 的功能受到 Toll 样受体 (TLR) 引发的信号的高度调控。因此，调节 TLR 信号通路的细胞内分子对于调节宿主免疫反应至关重要。我们最近发现 A20 是一种泛素修饰酶，可限制 Toll 样受体 (TLR) 诱导的信号。我们正在研究 DC 中 A20 表达在调节免疫稳态和反应中的作用。为了实现这一目标，我们培育了两种基因靶向小鼠品系：一种破坏所有细胞中的 A20 基因，另一种将 A20 的“floxed”等位基因靶向内源性 A20 基因座。我们将“floxed”小鼠与一种新的 CD11-Cre 转基因小鼠系杂交，该小鼠系在 DC 中以高选择性和特异性表达 Cre 酶。因此，这些复合 A20flox/flox CD11c-Cre 小鼠将为我们提供一种非常强大且新颖的试剂，用于研究 DC 中 A20 表达在调节免疫稳态和免疫反应中的作用。我们建议使用这些 A20flox/flox CD11-cre 复合小鼠以及源自这两种小鼠品系的细胞来确定 DC 中的 A20 表达如何调节其在体外和体内对 TLR 配体的反应。第一个目标将侧重于了解 A20 在调节 DC 中 TLR 信号传导中的作用的细胞生物学和生物化学，并将阐明受 A20 调节的 DC 的细胞自主功能。第二个目标将利用 A20flox/flox CD11c-Cre 小鼠来确定 DC 中的 A20 表达如何调节其体内稳态和激活。第二个目标还将探讨 DC 中的 A20 表达如何调节外周耐受和免疫反应。公共健康相关性：该项目重点研究一种称为 A20 的有效内源性抗炎蛋白，以及它如何调节体内树突状细胞功能。由于树突状细胞是免疫反应的强大调节剂，并且 A20 在调节树突状细胞功能中发挥着关键作用，因此了解 A20 如何调节树突状细胞和免疫反应可以显着促进炎症性疾病疗法的开发，从而对公众健康产生重大益处。