Genetic analysis of innate immunity using C. elegans

使用秀丽隐杆线虫进行先天免疫的遗传分析

基本信息

批准号：
7279917
负责人：
Alejandro Aballay
金额：
$ 26.28万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-30 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7279917
关键词：
Apoptosis Apoptotic Bacterial Infections Bioinformatics Caenorhabditis elegans Cell Death Cells Exhibits Genes Genetic Genetic Techniques Genomics Goals Gram-Positive Bacteria Heat shock proteins Heat-Shock Proteins 90 Homologous Gene Host Defense Immune response Infection Learning Libraries Lipopolysaccharides Mammals Mediating Methodology Missense Mutation Mitogen-Activated Protein Kinases Molecular Molecular Profiling Mus Natural Immunity Nematoda Pathway interactions Pattern Play Predisposition Publications Publishing RNA Interference Reporter Research Personnel Role Salmonella enterica Signal Pathway System Testing Tissues Translating Virulence Factors Work base comparative defense response design functional genomics genetic analysis human MAPK14 protein insight killings mutant pathogen positional cloning programs receptor research study response

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to provide insights into the molecular mechanism of innate immunity. Forward and reverse genetic studies have led to the identification of C. elegans signaling pathways that are required for defense response in both nematodes and mammals, suggesting that despite the vast evolutionary gulf between nematodes and mammals, some of the underlying mechanisms of defense response may be similar. We have used a set of C. elegans mutants hypersusceptible to pathogens to define a defense response pathway that involves the CED-3 programmed cell death pathway and the C. elegans homolog of the mammalian p38 mitogen-activated protein kinase (MAPK) encoded by the pmk-1 gene. Also, lipopolysaccharide (LPS) was found to act as a pathogen-associated molecular pattern (PAMP) that triggers a CED/MAPK-dependent programmed cell death in C. elegans. Our preliminary results indicate that ced-1 worms are hypersusceptible to S. enterica, ced-1, that is required for efficient clearance of apoptotic cells and cooperates with ced-3 to promote cell death, encodes a homolog of the mammalian CD91 receptor which has been involved in the elicitation of innate and adaptive immune responses by recognizing heat shock proteins, including HSP90. In addition, to establish parallels between innate immunity in mammals and nematodes, we have studied the expression profile of mice exposed to LPS to identify PAMPresponsive genes and the role of the C. elegans homologs in defense response was analyzed. Three C. elegans mutants in LPS-responsive genes were tested for their susceptibility to bacterial pathogens and two of them were found to be deficient in defense response. One of the mutants is daf-21(p673), which has a missense mutation in the C. elegans homolog of mammalian hsp90. First, we propose to continue our preliminary studies involving ced genes to determine the mechanisms by which these genes trigger defense response and the relationship between cell death and susceptibility to pathogens. We will also further characterize the role of DAF-21/HSP90 in defense-response and dissect its interaction with the CED-1/CD91 pathway. Finally, we propose to use bioinformatics and functional and comparative genomics to identify and characterize defense-related genes required for innate immunity in both nematodes and mammals.

描述（由申请人提供）：该提案的目的是提供有关先天免疫分子机制的见解。前进和反向遗传研究导致了秀丽隐杆线虫信号传导途径的鉴定，这些信号通路是线虫和哺乳动物中国防反应所必需的，这表明尽管线虫和哺乳动物之间存在巨大的进化湾，但防御反应的某些潜在机制可能是相似的。我们已经使用了一组可用于病原体的秀丽隐杆线虫突变体过度敏感，以定义涉及CED-3程序性细胞死亡途径的防御反应途径，而C. e秀拉属秀丽隐杆线虫同源于由哺乳动物p38有丝分裂原激活的蛋白激酶（MAPK）编码的哺乳动物p38 p38 p38。 PMK-1基因。同样，发现脂多糖（LPS）充当病原体相关的分子模式（PAMP），可触发秀丽隐杆线虫中CED/MAPK依赖的程序性细胞死亡。我们的初步结果表明，Ced-1蠕虫是肠链球菌CED-1的过度敏感，这是有效清除凋亡细胞并与Ced-3合作以促进细胞死亡所必需的，它编码具有哺乳动物CD91受体的同源物，该同源物具有具有的同源物，该受体具有具有通过识别包括HSP90在内的热休克蛋白来参与启发先天和适应性免疫反应。此外，为了在哺乳动物和线虫中的先天免疫之间建立相似之处，我们研究了暴露于LPS的小鼠的表达谱以鉴定调子基因，以及秀丽隐杆线虫同源物在防御反应中的作用。测试了LPS响应基因中的三个秀丽隐杆线虫突变体对细菌病原体的敏感性测试，其中两个被发现缺乏防御反应。其中一个突变体是DAF-21（P673），该突变体在哺乳动物HSP90的C.秀丽隐杆线虫同源物中具有错义突变。首先，我们建议继续我们的初步研究，涉及CED基因，以确定这些基因触发防御反应以及细胞死亡与病原体易感性之间的关系的机制。我们还将进一步表征DAF-21/HSP90在防御反应中的作用，并剖析其与CED-1/CD91途径的相互作用。最后，我们建议使用生物信息学以及功能和比较基因组学来识别和表征线虫和哺乳动物先天免疫所需的防御相关基因。