CANDIDA ADHERENCE & PENETRATION OF VASCULAR ENDOTHELIUM

对于了解传播性念珠菌的发病机制至关重要- 分离是阐明念珠菌从细胞中逃逸的事件。 血管内隔室及其穿透血管壁 在目标器官的入侵。 这个过程发生在一个环境中， 富含循环的体液和细胞成分， 血管内皮细胞的边界。 在上一次赠款期间 期间，利用血管室-内皮细胞模型 由人血管内皮细胞组成的细胞界面， 组织培养中，我们发现了配体依赖的证据， 念珠菌粘附内皮细胞的相互作用， 多克隆抗血清阻断粘附，并显示，对于 白细胞对内皮细胞的保护作用， 念珠菌组织渗透。 此外，我们还开发了一种ELISA 为了定量内皮细胞粘附，淘析器系统 用于增加监测中性粒细胞的额外维度， 内皮细胞分泌生理反应和方法 专注于念珠菌生理学的各个方面，如细胞壁 组成，细胞形态和分泌产物可能是 在内皮细胞粘附和渗透中起重要作用。 利用这些背景知识和我们开发的新方法 在上一个资助期间，我们将描述 粘附素配体的免疫沉淀 粘附素与阻断粘附的抗血清和与 生物化学提取的细胞壁成分，阐明 念珠菌穿透内皮细胞的机制 浅蓝菌素耐药、芽管阴性和蛋白酶阴性 突变体，确定中性粒细胞保护 内皮免受念珠菌损伤，表征趋化因子 内皮细胞分泌，并确定内皮细胞是否 增强循环嗜酸性假丝酵母菌的吞噬和杀伤作用 通过分泌细胞因子和/或与细胞因子的连续接触， 中性粒细胞

Critical to understanding the pathogenesis of disseminated candi- diasis is elucidation of the events of escape of Candida from the intravascular compartment and its penetration through vessel walls during invasion of target organs. This process occurs in a milieu rich in circulating humoral and cellular elements and across the boundary of the vascular endothelial cells. During the last grant period, utilizing a model of the vascular compartment-endothelial cell interface consisting of human vascular endothelial cells in tissue culture, we found evidence for a ligand-dependent interaction in adherence of Candida to endothelium, made a polyclonal antiserum that blocks adherence, and showed, for the first time, a protective role of leukocytes for endothelium during Candida tissue penetration. Additionally, we developed an ELISA for quantifying endothelial cell adherence, an elutriator system for adding additional dimensions to monitoring neutrophil and endothelial cell secretory physiological responses, and methods to focus on facets of Candida physiology such as cell wall composition, cell morphology, and secretory products likely to be important in adherence and penetration of endothelium. Using this background knowledge and the newer methods we developed during the previous funding period, we will characterize the nature of the adhesin ligand using immunoprecipitation of cell wall adhesins with antiserum that blocks adherence and with biochemically extracted cell wall constituents, elucidate mechanisms of Candida penetration of endothelium with selected cerulenin resistant, germ tube negative, and protease negative mutants, determine mechanisms by which neutrophils protect endothelium from Candida damage, characterize chemotactic factors secreted by endothelium, and determine whether endothelial cells enhance circulating neutrophil-Candida phagocytosis and killing through secretion of cytokines and/or contiguous contact with neutrophils.