CANDIDA ADHERENCE & PENETRATION OF VASCULAR ENDOTHELIUM

念珠菌依从性

• 批准号: 3129449
• 负责人: John E Edwards
• 金额: $ 11.06万
• 依托单位: LOS ANGELES COUNTY HARBOR-UCLA MED CTR
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3129449
• 关键词: Candida; blocking antibody; candidiasis; cell adhesion; cell cell interaction; cell wall; cellular pathology; chemoattractants; complement receptor; electrophoresis; enzyme linked immunosorbent assay; fungal antigens; glycoproteins; host organism interaction; human tissue; immunochemistry; laboratory rabbit; membrane activity; membrane proteins; membrane structure; microorganism immunology; mutant; neutrophil; phagocytosis; protein structure function; radioassay; surface antigens; tissue /cell culture; vascular endothelium

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.

对于了解播散性念珠菌的发病机制至关重要 diasis 是对念珠菌从 血管内室及其对血管壁的渗透 在侵袭靶器官时。 这个过程发生在一个环境中 富含循环体液和细胞元素并遍布全身 血管内皮细胞的边界。 上次拨款期间 期间，利用血管内皮细胞模型 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性的证据 念珠菌与内皮细胞粘附的相互作用， 多克隆抗血清可阻止粘附，并表明，对于 首次发现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 用于量化内皮细胞粘附，淘析系统 用于增加额外的维度来监测中性粒细胞和 内皮细胞分泌生理反应及其方法 关注念珠菌生理学的各个方面，例如细胞壁 组成、细胞形态和分泌产物可能 对内皮的粘附和渗透很重要。 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，我们将描述性质 使用细胞壁免疫沉淀法测定粘附素配体 具有阻止粘附的抗血清的粘附素和 生化提取细胞壁成分，阐明 念珠菌穿透内皮细胞的机制 浅蓝菌素抗性、芽管阴性、蛋白酶阴性 突变体，确定中性粒细胞保护的机制 念珠菌损伤的内皮细胞，表征趋化因子 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-念珠菌的吞噬和杀灭作用 通过分泌细胞因子和/或与 中性粒细胞。