Molecular and Immunologic Analysis of the Pathobiology of Human Anthrax

描述(由申请人提供)：在这份续签申请中，我们建议继续我们的多学科方法来研究人类中的炭疽杆菌感染。我们认为炭疽杆菌是作为生物恐怖分子对我们社会的最大威胁之一，有别于将其用作生物武器。炭疽杆菌作为恐怖武器令人担忧，原因有几个：1)孢子寿命长，在恶劣环境中稳定；2)疫苗质量差，效力可疑，注射程序繁琐，不提供给平民；3)这种疾病没有有效的治疗方法；4)一种罕见的外来疾病造成少量死亡，足以在我们的人口中造成恐惧。事实上，这正是2001年美国邮政服务中心遇袭后发生的事情。我们的应用程序建立在以前资助的U19基础上，用于研究人类和非人类灵长类动物对炭疽杆菌感染和免疫的反应。在第一次赠款期间，我们建立了一个拥有非凡的物质和智力资产以及一流的基础设施的组织。我们组建的团队富有成效，互动性很强，在免疫学方面拥有令人敬畏的背景。我们的团队包括四个不同机构的教职员工、研究员、技术人员、兽医人员和其他支持人员：俄克拉荷马医学研究基金会、俄克拉荷马大学、现在的芝加哥大学和波士顿大学。申请包括五个科学项目、三个技术项目和两个核心设施(以及行政和教育核心)。我们从各个方面着手解决这个问题：主要在内部开发的强大的实验模型，疫苗遗传学和疫苗效力的新方法，几个宿主反应问题，外毒素的研究，以及两种新的令人兴奋的治疗方法。总体而言，我们相信，四年的供资使我们处于具有竞争力的地位，能够利用我们第一个周期的调查结果，在未来五年取得实质性进展。 相关性(见说明)：炭疽杆菌是生物恐怖主义高度相关和重要的病原体。虽然疫苗是提供给军事人员的，但不是给普通公民的，其效力非常值得怀疑。人们对炭疽杆菌在人类中的感染知之甚少，很大程度上是因为炭疽病只是一种恐怖武器，自然的人类感染很少发生。 项目1：炭疽病期间水肿性毒素抑制免疫反应(Ballard，J) 项目1描述(由申请人提供)：由于炭疽芽孢杆菌能够避免宿主免疫系统的清除，炭疽病从最初的感染发展为严重的系统性疾病。炭疽毒素由保护性抗原(PA)、水肿因子(EF)和致死因子(LF)组成，是导致疾病的主要因素，因为该毒素抑制免疫细胞功能。因此，对炭疽毒素作用机制的深入了解为理解炭疽杆菌的发病机制提供了必要的关键信息。本项目旨在阐明水肿型毒素(ET：PA+EF)对先天性免疫反应的影响，并确定ET如何与致死性毒素(LT：PA+LF)结合来完成这一过程。在PA转位到细胞内后，EF作为腺苷环化酶发挥作用，并产生高水平的cAMP。在最近的研究中，我们发现ET激活糖原合成酶-3|3(GSK-3(3))，导致β-连环蛋白失活和3-连环蛋白协同转录调节丧失。这些研究的目的是阐明ET对免疫细胞功能的影响以及这一过程对人肺泡巨噬细胞和外周血单核细胞的影响，并确定ET和LT对这些细胞的联合作用。本项目的具体目标是：特定目标1：我们将描述在吸入性炭疽病早期阶段ET诱导的炎症反应和细胞内信号的变化，这些变化是导致吸入性炭疽病早期关键免疫抑制的原因。 相关性(见说明)：炭疽病是对美国人口健康的主要威胁，因为生物恐怖分子可能邪恶地使用炭疽杆菌。尽管有各种各样的治疗选择，炭疽病的死亡率很高。因此，了解炭疽杆菌毒力的基本机制仍然是至关重要的。因此，拟议的研究具有很高的相关性，因为这项工作的发现将有助于更好地了解疾病期间的免疫抑制，并确定预防和治疗的新途径。

DESCRIPTION (provided by applicant): In this renewal application we propose to continue our multidisciplinary approach to the study of Bacillus anthracis infections in humans. We view B. anthracis as one of the most significant threats to our society as a bioterrorist agent, distinct from its use as a biological weapon. B. anthracis is alarming as a weapon of terror for several reasons: 1) the spore is long-lived and stable in harsh environments; 2) the vaccine is poor, of questionable efficacy, has a cumbersome injection schedule, and is not provided to civilians; 3) there are no effective treatments for the disease; 4) a small number of deaths from a rare and thus exotic disease is sufficient to cause terror in our population. Indeed, this is exactly what happened after the US Postal Service Center attacks in 2001. Our application builds on a previously funded U19 to study the human and non-human primate response to B. anthracis infections and immunizations. During the first grant period, we built an organization with remarkable physical and intellectual assets and a superb infrastructure. The group we assembled is productive, highly interactive and has a formidable background in immunology. Our group includes faculty, fellows, technicians, veterinary staff and other support staff at four different institutions: The Oklahoma Medical Research Foundation, The University of Oklahoma, and now The University of Chicago, and Boston University. The application contains five Scientific Projects, three Technical Projects and two Core Facilities (along with Administrative and Educational cores). We attack the problem from every direction: powerful experimental models largely developed in house, a novel approach to vaccine genetics and vaccine efficacy, several host-response issues, studies of the exotoxins, and two novel and exciting therapeutic approaches. Overall, we are confident that the four years of funding puts us in a competitive position to exploit the findings of our first cycle to make substantive advances in the next five years. RELEVANCE (See instructions): B. anthracis is highly relevant and important agent of bioterrorism. Although a vaccine is provided to military personnel, it is not given to private citizens and is of highly questionable efficacy. There is very little understood of B. anthracis infections in humans, largely because anthrax is only a weapon of terror and natural human infections occur rarely. PROJECT 1: Edema Toxin Suppression of Immune Responses During Anthrax Disease (Ballard, J) PROJECT 1 DESCRIPTION (provided by applicant): Anthrax disease progresses from initial infection to serious systemic illness due to the ability of Bacillus anthracis to avoid clearance by the host immune system. Anthrax toxin, composed of protective antigen (PA), edema factor (EF), and lethal factor (LF), is a major contributing factor to disease as the toxin suppresses immune cell function. Thus, insights into the mechanism of action for anthrax toxin provides critical information necessary for understanding the pathogenesis of B. anthracis. In the current project experiments are designed to elucidate the effects of edema toxin (ET: PA plus EF) on innate immune responses, and determine how ET combines with lethal toxin (LT: PA plus LF) to accomplish this process. After translocation into the cell by PA, EF functions as an adenylate cyclase and generates high levels of cAMP. In recent studies we have discovered that ET activates glycogen synthase kinase-3|3 (GSK- 3(3) leading to inactivation p-catenin and loss in (3-catenin cotranscriptional regulation. The goal of these studies are now to elucidate the impact ET-mediated disruption immune cell function and the effects of this process on human alveolar macrophages and peripheral blood mononuclear cells, as well as determine the combined effects of ET and LT on these cells. The specific aims of this project are: Specific Aim 1: We will characterize the ET-induced changes in inflammatory responses and intracellular signaling that account for critical immunosuppression during early stages of inhalational anthrax Specific Aim 2: We will characterize the ET-induced changes in inflammatory responses and intracel signaling that account for critical immunosuppression during late stages of inhalational anthrax Specific Aim 3: We will characterize the combined effects of ET and LT on immunosuppression during both early and late stages of anthrax disease. RELEVANCE (See instructions): Anthrax is a major threat to the health of US population, because of the potential nefarious use of B. anthracis by bioterrorist. Anthrax has a high mortality rate, despite various treatment options. Thus it continues to be essential to understand the basic mechanisms of B. anthracis virulence. For this reason the proposed studies are highly relevant as findings from this work will help better understand immune suppression during disease and identify new avenues for prophylactic and therapeutic treatments.