CANDIDA ADHERENCE & PENETRATION OF VASCULAR ENDOTHELIUM

对了解播散性心绞痛的发病机制至关重要 假丝酵母菌逃逸事件是指假丝酵母菌 血管内室及其对血管壁的穿透 在入侵目标器官期间。此过程在一个环境中进行 富含循环中的体液和细胞元素， 血管内皮细胞的边界。在最后一次拨款期间 期间，利用血管腔内皮细胞模型 由人血管内皮细胞组成的细胞界面 组织培养，我们发现了配体依赖的证据 念珠菌与内皮细胞黏附的相互作用 阻止黏附的多克隆抗血清，并显示，对于 首次发现白细胞对血管内皮细胞的保护作用 假丝酵母菌组织穿透。此外，我们还开发了一种酶联免疫吸附试验 为了量化内皮细胞的黏附，一种淋洗系统 为了增加监测中性粒细胞和 内皮细胞分泌生理反应及其方法 关注假丝酵母菌生理方面，如细胞壁 成分、细胞形态和可能的分泌产物 对内皮细胞的黏附和穿透很重要。 利用这一背景知识和我们开发的较新方法 在上一个资助期内，我们将描述 用细胞壁免疫沉淀法测定粘附素配体 粘附素与阻止粘连的抗血清和与 生物化学提取的细胞壁成分，阐明 白念珠菌穿透血管内皮细胞的机制 蓝菌素耐药，芽管阴性，蛋白酶阴性 突变体，决定中性粒细胞保护机制 念珠菌损伤的内皮细胞、趋化因子的特征 由内皮细胞分泌，并确定内皮细胞是否 增强循环中性粒细胞-念珠菌的吞噬和杀伤作用 通过分泌细胞因子和/或与 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.