Viral Immunity in Drosophila

果蝇的病毒免疫

• 批准号: 7010727
• 负责人: NORBERT PERRIMON
• 金额: $ 33.1万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7010727
• 关键词: Drosophilidae; Picornaviridae; arthropod genetics; biological signal transduction; double stranded RNA; endocytosis; functional /structural genomics; high throughput technology; host organism interaction; microarray technology; microorganism immunology; tissue /cell culture; toll like receptor; virus infection mechanism; virus replication

DESCRIPTION (provided by applicant): Studies on Drosophila innate immunity have highlighted the evolutionary conservation of response mechanisms with regards to bacterial and fungal infections. Indeed the role of the Toll receptors in mammalian immunity originated from studies on immunity in flies. In this proposal we aim to dissect the mechanisms of anti-viral innate immunity in Drosophila. In mammals, viruses trigger both acquired and innate immune responses. While the role of the innate immune system in anti-viral immunity is well established, many aspects of this response are still poorly understood. Because Drosophila has a mechanism to fight viruses, we propose to use this model organism, and take an unbiased genetic approach to identify the pathways that are required for viral immunity. Our hypothesis is that by studying immunity to viral pathogens in Drosophila, which does not have an acquired immune system, we will be able to identify conserved innate mechanisms by which organisms fight viruses. Viruses are a class of obligate intracellular pathogens that require host factors for their replication and survival. We have established both in vivo and in vitro assays to analyze the response of adult flies and cells to a Drosophila C picornavirus. We will use the powerful tools of genetics and functional genomics available in Drosophila to identify host factors involved in viral replication and immune protection to this RNA virus. Further, we will analyze the transcriptional response of host cells to viral infection to identify genes that are involved in the viral response. Using a robust tissue culture assay, which is based on the ability of the Drosophila C picornavirus to kill cells, we will conduct a highthroughput RNA interference screen to identify all proteins present in the Drosophila genome that when absent prevent viral infection. This assay will also be used to perform a chemical screen to identify small molecules that protect cells from viral infection. Because our insect virus system is sensitive to drugs that are used against human viruses, our studies may lead to the identification of novel antiviral compounds that may prove effective in mammalian cells.

描述（由申请人提供）：对果蝇先天免疫的研究强调了细菌和真菌感染反应机制的进化保守性。事实上，Toll 受体在哺乳动物免疫中的作用源于对果蝇免疫的研究。在本提案中，我们的目的是剖析果蝇抗病毒先天免疫的机制。在哺乳动物中，病毒会触发获得性和先天性免疫反应。虽然先天免疫系统在抗病毒免疫中的作用已得到充分证实，但这种反应的许多方面仍然知之甚少。由于果蝇具有对抗病毒的机制，我们建议使用这种模型生物，并采取公正的遗传方法来识别病毒免疫所需的途径。我们的假设是，通过研究果蝇（不具有获得性免疫系统）对病毒病原体的免疫力，我们将能够识别生物体对抗病毒的保守先天机制。病毒是一类专性细胞内病原体，其复制和生存需要宿主因子。我们已经建立了体内和体外试验来分析成年果蝇和细胞对果蝇 C 小核糖核酸病毒的反应。我们将利用果蝇中可用的强大的遗传学和功能基因组学工具来识别参与病毒复制和对该 RNA 病毒的免疫保护的宿主因子。此外，我们将分析宿主细胞对病毒感染的转录反应，以确定参与病毒反应的基因。使用基于果蝇 C 小核糖核酸病毒杀死细胞的能力的强大组织培养测定，我们将进行高通量 RNA 干扰筛选，以识别果蝇基因组中存在的所有蛋白质，这些蛋白质在缺乏时可防止病毒感染。该测定还将用于进行化学筛选，以识别保护细胞免受病毒感染的小分子。由于我们的昆虫病毒系统对用于对抗人类病毒的药物敏感，因此我们的研究可能会导致鉴定出可能对哺乳动物细胞有效的新型抗病毒化合物。