Hemocyte Transcriptome and Immunity in Aedes aegypti

埃及伊蚊的血细胞转录组和免疫

• 批准号: 7021822
• 负责人: BRUCE MARTIN CHRISTENSEN
• 金额: $ 36.38万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-15 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021822
• 关键词: Aedes; Klebsiella pneumoniae; Micrococcus; RNA interference; Salmonella typhimurium; Staphylococcus; arthropod genetics; blood cells; cellular immunity; disease vectors; double stranded RNA; gene expression profiling; genetic transcription; hemolymph; high throughput technology; host organism interaction; humoral immunity; immune response; immunogenetics; laboratory rabbit; microarray technology; microorganism culture; molecular biology information system; oligonucleotides; phagocytosis; phenotype; synthetic peptide

DESCRIPTION (provided by applicant): The mosquito innate immune response is a major factor governing the interaction between vector and pathogen; consequently, it is a primary determinant of vector competence. Innate immunity in mosquitoes and other insects employs both cellular and humoral components in response to invading pathogens, but studies of the humoral aspects of innate immunity have dominated, with a primary emphasis on the identification of antimicrobial peptides (AMPs) and the signaling pathways involved in their production. Far less emphasis has been placed on hemocytes and cellular immune responses, but recent data clearly verify that cellular responses likely play a critical role in clearing microbial pathogens by phagocytosis, melanization, or by both processes in combination. There is a void in the data required to accurately define regulatory processes, hemocyte-produced effector molecules, and the role these cells and cell products play in innate immunity. It also is clear that AMPs are significant factors in innate immune responses against invading microbes, but it is not known how cellular and AMP-based humoral responses function in concert to clear microbial infections. We believe that a reductionist approach, that studies one gene, or a small set of genes, in relation to a phenotype, has limitations for determining the complex interrelationships that undoubtedly exist in mosquito innate immune responses. Therefore, in order to initate more holistic experimental approaches for studying innate immunity in mosquitoes, we developed a large expressed sequence tag (EST) data set from Aedes aegypti immune-activated hemocytes and complementary oligonucleotide-based microarrays as substrates to evaluate mosquito cellular, as well as humoral, immune responses. The general hypothesis for the research proposed herein is that mosquito innate immune responses are a highly complex network of processes that employ the distinct, but not mutually exclusive, responses of phagocytosis and melanization, in concert with AMPs, to clear bacteria infections from the hemolymph of infected mosquitoes. To test this hypothesis, we will use the mosquito Ae. aegypti and a number of bacteria species in (1) high-throughput molecular approaches to profile gene transcription and to identify distinct patterns underlying the anti-bacterial responses of phagocytosis and melanization, (2) utilize microarray analyses to assess transcription profile changes in relation to different bacterial species (that vary in their pathogenicity and ability to elicit a response) in order to better dissect mechanisms influencing cellular immunity, and (3) employ microarray analyses, synthetic peptides, RNAi methodologies, and phenotype evaluations to better define the role AMPs, and select unknown genes, play in innate immunity. All data resulting from microarray analyses, including experimental protocols used, will be made publicly available on the interactive website that contains our hemocyte EST data.

描述（由申请人提供）：蚊子先天免疫反应是控制载体与病原体之间相互作用的主要因素。因此，它是向量能力的主要决定因素。蚊子和其他昆虫的先天免疫力对入侵病原体采用了细胞和体液成分，但是对先天免疫的体液方面的研究已经占主导地位，主要强调了抗菌肽（AMP）的鉴定以及涉及其生产中的信号通路。对血细胞和细胞免疫反应的重视程度要少得多，但是最近的数据清楚地证明了细胞反应在通过吞噬细胞增多症，黑色素化或两种过程中的结合过程中可能起关键作用。准确定义调节过程，血细胞产生的效应子分子以及这些细胞和细胞产物在先天免疫中所起的作用所需的数据中存在一个空隙。同样很明显，AMP是针对入侵微生物的先天免疫反应的重要因素，但尚不知道基于细胞和AMP的体液反应如何协同起作用以清除微生物感染。我们认为，一种还原主义的方法，即研究一个基因或与表型有关的一小部分基因具有确定蚊子先天免疫反应中无疑存在的复杂相互关系的局限性。 Therefore, in order to initate more holistic experimental approaches for studying innate immunity in mosquitoes, we developed a large expressed sequence tag (EST) data set from Aedes aegypti immune-activated hemocytes and complementary oligonucleotide-based microarrays as substrates to evaluate mosquito cellular, as well as humoral, immune responses.本文提出的研究的一般假设是，蚊子先天免疫反应是一种高度复杂的过程网络，它采用了吞噬作用和黑质的独特但不排斥的反应，与AMPS协同，以清除感染蚊子血液抑制细菌感染的细菌感染。为了检验这一假设，我们将使用蚊子AE。 （1）（1）高通量分子方法的埃及和许多细菌物种，用于剖析基因转录，并​​确定吞噬作用和黑色素化的抗细菌反应的不同模式（（2）利用微阵列分析来评估其与其他细菌的变化（效率）的变化（效果效率）（该效率及其在效果中）的效果（效率）（该效率）（该物种效果均具有效率（该物种），并在该物种中的变化（该病态依赖性）（该效率）（该方法）的病态依赖性（效率）（该物种）的病态依赖性（效果）（影响细胞免疫，（3）采用微阵列分析，合成肽，RNAi方法和表型评估，以更好地定义角色AMP并选择未知基因，以天生的免疫力发挥作用。由微阵列分析产生的所有数据（包括所使用的实验协议）将在包含我们的Hemocyte EST数据的交互式网站上公开提供。