Innate immunity to adenovirus vectors

对腺病毒载体的先天免疫

• 批准号: 6954293
• 负责人: Dmitry Shayakhmetov
• 金额: $ 25.77万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954293
• 关键词: Adenoviridae; capsid; cell type; genetically modified animals; green fluorescent proteins; host organism interaction; immune response; inflammation; integrins; laboratory mouse; low density lipoprotein receptor; mass spectrometry; microorganism immunology; polymerase chain reaction; protein protein interaction; receptor binding; receptor expression; site directed mutagenesis; transfection /expression vector; virus diseases; virus infection mechanism; virus receptors

DESCRIPTION (provided by applicant): This proposal seeks to identify the exact step(s) of adenovirus (Ad) infection that is (are) responsible for the initiation of an anti-Ad acute inflammatory response upon systemic virus application. Over the last two decades numerous Ad-based vectors have been developed for gene therapy applications and many are currently being tested in clinical trials. Most recently, interest in Ad has further expanded due to its potential as a vector for vaccination against life threatening infectious agents such as anthrax. While natural infections with Ad are largely harmless to humans, intravenous Ad administration may result in a severe inflammatory response, which can lead to fatal outcomes. It is currently recognized that the initiation of this acute systemic inflammation depends on interactions of the Ad capsid with host cells. Despite significant knowledge regarding Ad interactions with cells in vitro, the molecular mechanisms governing Ad bio-distribution, hepatic tropism and toxicity in vivo remain poorly understood. Recently, we identified a novel blood factor-dependent pathway of Ad liver cell infection in vivo. This finding explained the observed Ad biodistribution in animals after systemic application and is fundamental for the development of novel strategies to modify both Ad tropism in vivo and virus-associated toxicity. In mouse models, we will analyze the innate immune response to intravenously applied capsid-modified Ad mutants deficient in their ability to undergo the initial steps of virus infection. Our specific aims are to analyze the role of Ad interactions 1) with primary attachment receptors; 2) with integrins, which facilitate Ad initialization; or 3) with cellular factors upon virus escape from endosomes, in the initiation of an anti-Ad inflammatory response. Based on the data obtained, in our fourth specific aim we will construct ah Ad vector ablated for binding to known virus receptors and capable of infecting cells via an alternative receptor, LDLR. By applying this vector in mice and evaluating its toxicity we will test the hypothesis that modification of the Ad capsid represents a feasible approach to reduce acute Ad-mediated inflammation while preserving virus gene transfer ability upon systemic application. These studies will significantly improve our understanding of fundamental mechanisms of the host defense against viral pathogens and may ultimately lead to the development of safe and efficient Ad vectors for the therapy of a wide range of inborn and acquired human diseases.

描述（由申请人提供）：该提案旨在确定（AD）感染的确切步骤，该步骤是（）负责在全身病毒应用时启动抗AD急性炎症反应的原因。在过去的二十年中，已经为基因治疗应用开发了许多基于AD的载体，目前正在临床试验中进行了测试。最近，由于它作为疫苗接种疫苗的潜力，对炭疽等感染者的疫苗接种潜力，因此对AD的兴趣进一步扩大。尽管AD的自然感染对人类无害，但静脉内给药可能会导致严重的炎症反应，这可能导致致命的结果。目前认识到，这种急性全身性炎症的启动取决于AD CAPSID与宿主细胞的相互作用。尽管有关AD与细胞体外相互作用的知识很高，但管理AD生物分布的分子机制，体内的肝湿度和毒性仍然知之甚少。最近，我们确定了体内AD肝细胞感染的新型血液因子依赖性途径。这一发现解释了全身应用后观察到的动物中观察到的AD生物分布，这对于开发了新的策略来改变体内AD tropism和病毒相关的毒性。在小鼠模型中，我们将分析对静脉施用的衣壳修饰的AD突变体的先天免疫反应，因为其经历病毒感染的初始步骤的能力不足。我们的具体目的是分析AD相互作用的作用1）与主要附着受体的作用； 2）与整合素一起，有助于AD初始化；或3）在抗AD炎症反应开始时，病毒后，病毒逃脱了细胞因子。基于获得的数据，在我们的第四个特定目的中，我们将构建用于结合已知病毒受体的AH AD载体，并能够通过替代受体LDLR感染细胞。通过将该载体应用于小鼠并评估其毒性，我们将检验以下假设：AD CAPSID的修饰代表了一种可行的方法，可减少急性AD介导的炎症，同时在全身应用下保留病毒基因转移能力。这些研究将显着提高我们对宿主防御病毒病原体的基本机制的理解，并最终可能导致开发安全有效的AD媒介，以治疗多种先天和获得的人类疾病。