AIDS OPPORTUNISTIC INFECTIONS--VIRULENCE OF C GLABRATA

描述（改编自申请人摘要）：在HIV阳性人群中，阴道和食管念珠菌病的第二大常见原因是光滑念珠菌。这部分是由于C的相对电阻。光滑到唑治疗。C. glabrata在ICU中也很普遍。C. glabrata与宿主的关系尚不清楚。本研究的目的是分析C.光滑念珠菌与寄主的关系，并对光滑念珠菌的毒力因子进行鉴定。光滑的分子和遗传操作在C. glabrata，并且由于C. glabrata是单倍体。研究人员已经产生了随机插入突变体，并开发了一种策略，在单个池中平行筛选多达96个突变体。使用这种策略，他已经确定了EPA1基因，这是一种介导与宿主细胞表面碳水化合物结合的粘附素。在这个提议中，他将描述这种粘附素的配体特异性，结构域结构和亚细胞定位。遗传和分子方法将被用来确定其他粘附素，最终的目标是确定大多数C。glabrata粘附素相关的宿主细胞的相互作用。在EPA1分析中开发的遗传工具允许对毒力进行广泛的遗传分析。研究者将在粘膜和系统性念珠菌病的小鼠模型中直接筛选无毒力突变体。许多二次筛选将鉴定那些涉及与特定宿主分子的离散相互作用的突变体。

DESCRIPTION (Adapted from the Applicant's Abstract): In the HIV- positive population, the second most frequent cause of vaginal and esophageal candidiasis is Candida glabrata. This is in part due to the relative resistance of C. glabrata to azole therapeutics. C. glabrata is also prevalent in ICUs. The interaction of C. glabrata with the host is not well understood. The objective of this proposal is to analyze the molecular interactions of C. glabrata with its host and to identify virulence factors in C. glabrata. Molecular and genetic manipulation is possible in C. glabrata, and is made easier by the fact that C. glabrata is haploid. The investigator has generated random insertion mutants and developed a strategy to screen up to 96 mutants in parallel in a single pool. Using this strategy, he has identified the EPA1 gene, an adhesin mediating binding to carbohydrates on the cell surface of the host. In this proposal, he will characterize this adhesin's ligand specificity, domain structure and sub-cellular localization. Genetic and molecular approaches will be used to identify other adhesins, with the eventual aim to characterize the majority of C. glabrata adhesins relevant to the host-cell interaction. The genetic tools developed in the analysis of EPA1 allow for a broad genetic analysis of virulence. The investigator will screen directly for avirulent mutants in murine models of mucosal and systemic candidiasis. A number of secondary screens will identify those mutants involved in discrete interactions with specific host molecules.