Discovery of New Cellular Targets of Anthrax Toxin

炭疽毒素新细胞靶点的发现

• 批准号: 6675112
• 负责人: ALINE BETANCOURT
• 金额: $ 22.28万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6675112
• 关键词: Bacillus anthracis; RNase protection assay; antibacterial agents; antigen antibody reaction; bacteria infection mechanism; bacterial toxins; bacterial vaccines; biological signal transduction; bioterrorism /chemical warfare; cellular immunity; cytokine; dendritic cells; enzyme linked immunosorbent assay; host organism interaction; immunoprecipitation; laboratory mouse; macrophage; microarray technology; mitogen activated protein kinase; molecular shape; northern blottings; pathologic process; polymerase chain reaction; proteomics; tissue /cell culture; toxicant interaction

DESCRIPTION (provided by applicant): The post- September 11 release of anthrax spores resulted in five civilian deaths, eighteen infections, and required that more than 30,000 individuals undergo prophylactic antibiotic therapy. This event also highlighted the need for a more thorough understanding of the pathogenesis of anthrax, and improved vaccines that would be appropriate for pre- or post-exposure immunization of civilian and military populations. The interaction of macrophages with one of the principal toxins produced by Bacillus anthracis, Lethal Toxin, has been a primary focus of laboratories studying the pathogenesis of this organism. However, we now know that dendritic cells are important sentinel cells distributed throughout the body that play a major role in detecting invading pathogens and in the initiation of the host immune response to foreign antigens. Our preliminary studies have shown that anthrax toxin affects cell signaling pathways and cytokine expression in toxin-treated dendritic cells. We will test the hypothesis that anthrax toxin targets several unknown but key molecular pathways within the dendritic cell that disarm its critical function in host-pathogen defense. The first specific aim is to determine the molecular changes within dendritic cells treated with anthrax toxin. We will use both gene microarray analysis and traditional cell signaling kinase assays to learn about both genomic and proteomic changes elicited by toxin treatment of these cells. The second aim is to determine the changes in cytokine expression profiles by dendritic cells treated with anthrax toxin. This will be measured by cytometric bead arrays, ELISA and RNase protection assays and is used here to reveal how the anthrax toxin-treated dendritic cell might affect other host cells and in particular those involved in host immune responses. This combined approach will significantly advance our understanding of B. anthracis pathogenesis in two ways: it will elucidate the dendritic cell's role in B. anthracis pathogenesis and it will lead to the identification of new cellular targets of anthrax toxin. Moreover, because of the critical role that dendritic cells play in initiating the host immune response, the proposed studies may lead to the development of improved vaccines against anthrax. This body of information will also serve as the foundation for new research areas to explore in future proposals.

描述（由申请人提供）：9月11日释放炭疽孢子的释放导致了5例平民死亡，十种感染​​，并要求30,000多人接受预防性抗生素疗法。 这项事件还强调了需要对炭疽病的发病机理进行更彻底的了解，并改善了适合于平民和军事人群的暴露前或暴露后免疫的疫苗。 巨噬细胞与炭疽芽孢杆菌（致命毒素）产生的主要毒素之一的相互作用一直是研究这种生物体发病机理的实验室的主要重点。 但是，我们现在知道树突状细胞是分布在整个人体中的重要哨兵细胞，在检测入侵病原体和宿主对外抗原的免疫反应的启动方面起着重要作用。 我们的初步研究表明，炭疽毒素会影响毒素处理的树突状细胞中细胞信号通路和细胞因子的表达。 我们将检验以下假设：炭疽毒素靶向树突状细胞中的几种未知但关键的分子途径，这些途径在宿主 - 病原体防御中解除了关键功能。 第一个具体目的是确定用炭疽毒素处理的树突状细胞内的分子变化。 我们将同时使用基因微阵列分析和传统的细胞信号激酶分析来了解这些细胞的毒素处理引起的基因组和蛋白质组学变化。 第二个目的是确定用炭疽毒素治疗的树突状细胞的细胞因子表达谱的变化。 这将通过细胞量珠阵列，ELISA和RNase保护分析来衡量，并在此用来揭示炭疽毒素处理的树突状细胞如何影响其他宿主细胞，尤其是参与宿主免疫反应的宿主细胞。 这种合并的方法将通过两种方式显着提高我们对炭疽芽孢杆菌发病机理的理解：它将阐明树突状细胞在炭疽芽孢杆菌发病机理中的作用，并将导致鉴定炭疽毒素的新细胞靶标。 此外，由于树突状细胞在启动宿主免疫反应中起着至关重要的作用，因此提出的研究可能导致对炭疽病的改善疫苗的发展。 这一信息还将为未来建议中探索的新研究领域提供基础。