Studies on Macrophage Resistance to Anthrax Lethal Toxin

巨噬细胞对炭疽致死毒素的抵抗力研究

• 批准号: 6804467
• 负责人: MOLLY A HUGHES
• 金额: $ 26.62万
• 依托单位: UNIVERSITY OF VIRGINIA
• 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 2007-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6804467
• 关键词: Bacillus anthracis; anthrax; anthrax toxin; bacterial cytopathogenic effect; bacterial proteins; bactericidal immunity; biological signal transduction; bioterrorism /chemical warfare; confocal scanning microscopy; cytokine; gene expression; genetic library; host organism interaction; immunofluorescence technique; immunoprecipitation; kinesin; laboratory mouse; macrophage; microarray technology; mitogen activated protein kinase; protein localization; protein protein interaction; protein transport; proteomics; virulence; yeast two hybrid system

DESCRIPTION (provided by applicant): Lethal toxin is a major virulence factor of Bacillus anthracis. Lethal toxin (LT) is comprised of two proteins, protective antigen (PA) and lethal factor (LF). Macrophages serve as the first line of defense against anthrax infection. There is a striking difference in susceptibility of mouse strains and the primary macrophages or macrophage cell lines derived from those strains to lethal, toxin-mediated effects. Certain mouse strains and their macrophages are sensitive to LT whereas other mouse strains and their macrophages are highly resistant to LT. The underlying mechanism of resistance remains unknown and is a subject of great interest in the anthrax field due to the potential usefulness of this information to development of therapeutics. The primary goal of this project is to investigate mechanisms of resistance to LT in macrophages. To address this goal, three specific aims are proposed: (1) Investigation of alterations in cellular trafficking and localization using immunofluorescence/confocal microscopy to examine localization of LF in sensitive versus resistant macrophages, (2) Investigation of the role of potential LF intracellular substrates in resistant macrophages, and (3) Analysis of differential microarray analyses to evaluate gene expression in LT-sensitive versus resistant macrophages with a focus on evaluating cytokine expression, MAPK kinase effector molecule expression, and expression of Kif1C, a recently described murine gene product associated with resistance to LT. Successful completion of these studies will yield a fundamental understanding of mechanisms of macrophage resistance to LT.

描述（申请人提供）：致死毒素是炭疽杆菌的主要毒力因子。 致死毒素（LT）由两种蛋白质组成：保护性抗原（PA）和致死因子（LF）。 巨噬细胞是抵抗炭疽感染的第一道防线。 小鼠品系和原代巨噬细胞或源自这些品系的巨噬细胞系对致命的毒素介导作用的敏感性存在显着差异。 某些小鼠品系及其巨噬细胞对 LT 敏感，而其他小鼠品系及其巨噬细胞对 LT 高度耐药。 耐药性的潜在机制仍然未知，并且由于该信息对治疗方法的开发具有潜在的用途，因此成为炭疽领域非常感兴趣的主题。 该项目的主要目标是研究巨噬细胞对 LT 的抵抗机制。 为了实现这一目标，提出了三个具体目标：(1) 使用免疫荧光/共聚焦显微镜研究细胞运输和定位的变化，以检查敏感巨噬细胞与耐药巨噬细胞中 LF 的定位，(2) 研究潜在 LF 细胞内底物的作用(3) 差异微阵列分析，以评估 LT 敏感型与耐药型巨噬细胞中的基因表达，重点是评估细胞因子表达、MAPK 激酶效应分子表达和Kif1C，最近描述的一种与 LT 抗性相关的小鼠基因产物。 成功完成这些研究将使人们对巨噬细胞对 LT 的抵抗机制有一个基本的了解。