CELL SURFACE DYNAMICS OF MANNOPROTEINS OF C. ALBICANS

白色念珠菌是一种机会性的二相性真菌，能够引起 人类中的毁灭性疾病。最近的观察表明， 电池的表面形貌是动态的，而不是静态的和可变的 表面成分的表达可能不仅仅发生在生长在 不同的条件，但在单个细胞的生命周期内。是这样的 动力学反映了细胞壁成分的合成机制和 它们的组装成一个功能结构，并提出了另一个 监管的层级。它们还影响到共生和致病 在生物体定殖时生物体与寄主的关系 正常宿主并逃脱正常防御以建立感染 被入侵的主机。 这项建议关注的是甘露糖蛋白，这些蛋白是研究最少的 墙的成分，并具有最大的序列多样性潜力。 甘露糖蛋白已被证明存在于细胞表面和两者 甘露聚糖和蛋白质被认为与真菌的粘连有关 组织。 这项研究将采取生产单抗的方法来 甘露糖蛋白的水合物和蛋白质部分的鉴定 曲面组件。这些抗体的反应性将被确定。 整个细胞使用间接免疫荧光。自然界和 将使用免疫印迹法检查抗原决定簇的身份 技术、抗原对各种试剂和其他试剂的敏感性 技巧。生长和静止状态下表位表达的程度 在28℃和37℃下培养的相酵母菌和芽管 在37摄氏度下培养将被确定。表达的方式， 决定因素的表达和性质的变化的动力学将是 审查模式，以开发监管的假设 表达和影响这种变化的机制。这个 抗体与表面成分发生反应的能力将在 真菌与人体黏附过程中宿主-寄生虫相互作用的研究 上皮细胞。这样的抗体将被用来选择突变体 表面表位表达的改变和这些突变体的特征 用于表面表达和组织粘连。

Candida albicans is an opportunistic dimorphic fungus capable of causing devastating disease in humans. Recent observations have suggested that the surface topography of the cell is dynamic rather than static and variable expression of surface components may occur not just on cells grown under different conditions but on single cells during their lifetime. Such dynamics reflect on mechanisms of synthesis of the cell wall components and their assembly into a functional structure and suggest an additional hierarchy of regulation. They also impact on the commensal and pathogenic relationship of the organism and the host as the organism colonizes the normal host and escapes normal defenses to establish infection in the compromised host. This proposal focuses on the mannoproteins which are the least studied of the wall constituents and have potential for greatest sequence diversity. Mannoprotein has been shown to be at the surface of the cell and both mannan and protein have been implicated in adherence of the fungus to tissue. The research will take the approach of producing monoclonal antibodies to both car bohydrate and protein portions of mannoproteins to identify surface components. The reactivity of these antibodies will be determined with whole cells using indirect immunofluorescence. The nature and identity of the antigenic determinant will be examined using immunoblotting techniques, sensitivity of the antigen to various reagents and other techniques. The extent of epitope expression by growing and stationary phase yeasts cultivated at 28 degrees C and 37 degrees C and germ tubes cultivated at 37 degrees C will be determined. The patterns of expression, kinetics of changes in expression and nature of the determinant will be examined for patterns to develop hypotheses for the regulation of expression and the mechanisms by which such changes are effected. The ability of antibodies to react with surface components will be exploited in studying host-parasite interactions in adherence of fungi to human epithelial cells. Such antibodies will be utilized to select for mutants altered in expression of surface epitopes and these mutants characterized for surface expression and tissue adherence.