CRYPTOCOCCAL INTERACTIONS WITH HOST CELLS

隐球菌与宿主细胞的相互作用

基本信息

批准号：
8420419
负责人：
Tamara L Doering
金额：
$ 42.6万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-01 至 2015-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Cryptococcus neoformans is an opportunistic pathogen that is responsible for life-threatening disease in patients with AIDS or other conditions of immunocompromise; extrapulmonary cryptococcal infection is an AIDS-defining illness. The long-term goal of our studies is to understand the pathogenesis of cryptococcosis. This application focuses on a fundamental part of this process, the interactions of this facultative intracellular pathogen with the phagocytic cells of its mammalian host. These interactions resolve with varying levels of damage to the host and yeast, but the mechanisms involved are poorly understood, limiting our ability to influence resolution in favor of the host. We propose to combine forward genetics, high-throughput screening assays, and focused follow-up studies to determine the cryptococcal factors that are required for this pathogen to successfully adhere to and enter host cells. Aim I is to identify mutants in these processes, using insertional mutagenesis and automated screening. Aim II is to determine the mechanisms by which selected gene products influence pathogen adherence and entry interactions. These studies will employ assays of cell adhesion, entry, and intracellular survival and growth; molecular techniques for sequence recovery, analysis, and gene deletion; and phenotypic assessment including morphological studies, biochemical analysis, and virulence assays. Results will be assessed using rigorous statistical methods and appropriate controls, and will be interpreted in the context of our expertise in the fundamental biology of Cryptococcus. The proposed studies will elucidate a crucial aspect of pathogenesis that determines the survival and dissemination of the organism and the host's ability to limit disease. This work will additionally open new areas for investigation of an important pathogen and develop experiment tools that can be applied to other areas of fungal biology.

描述（由申请人提供）：新型隐球菌是一种机会性病原体，可导致艾滋病或其他免疫功能低下患者罹患危及生命的疾病；肺外隐球菌感染是一种艾滋病定义的疾病。我们研究的长期目标是了解隐球菌病的发病机制。本申请重点关注该过程的基本部分，即兼性细胞内病原体与其哺乳动物宿主的吞噬细胞的相互作用。这些相互作用会对宿主和酵母造成不同程度的损害，但所涉及的机制知之甚少，这限制了我们影响有利于宿主的解决方案的能力。我们建议结合正向遗传学、高通量筛选分析和重点后续研究，以确定该病原体成功粘附并进入宿主细胞所需的隐球菌因子。我的目标是使用插入诱变和自动筛选来识别这些过程中的突变体。目标 II 是确定所选基因产物影响病原体粘附和进入相互作用的机制。这些研究将采用细胞粘附、进入以及细胞内存活和生长的分析；用于序列恢复、分析和基因删除的分子技术；表型评估，包括形态学研究、生化分析和毒力测定。结果将使用严格的统计方法和适当的控制进行评估，并将根据我们在隐球菌基础生物学方面的专业知识进行解释。拟议的研究将阐明决定生物体生存和传播以及宿主限制疾病能力的发病机制的一个关键方面。这项工作还将开辟重要病原体研究的新领域，并开发可应用于真菌生物学其他领域的实验工具。