CELL SURFACE DYNAMICS OF MANNOPROTEINS OF C ALBICANS

描述（改编自申请人的摘要）：白色念珠菌是一种 机会性二型真菌，能够引起毁灭性疾病， 人类 最近的研究表明，细胞的表面形貌是 动态的而不是静态的，变化不仅发生在生长的细胞上， 在不同的条件下，但在没有细胞的情况下， 师. 这种动态反映了合成和调节机制 细胞壁组件及其组装成功能结构。 它们还影响生物体与宿主之间的相互作用， 健康和疾病，因为许多这些相互作用是在介导的 细胞表面 该建议集中在甘露糖蛋白和蛋白质上， 这是最不明确的成分，有可能 多样性。 这些组件已作为一个类显示 并与真菌对宿主组织的粘附有关 和宿主蛋白质的结合。 这项研究将采取生产单克隆抗体的方法， 这些成分的碳水化合物和蛋白质部分， 表面部分。 抗体也将产生的成分 与宿主蛋白质结合和真菌形态发生有关。 的 这些抗体的反应性将用全细胞测定， 间接免疫荧光法 抗原的性质和身份 将使用免疫印迹技术检查决定簇，灵敏度 抗原的各种试剂和其他技术。 的程度 通过生长期和稳定期酵母和芽管表达表位 将被确定。 由于该生物体暴露于多种抗体 和宿主中的宿主蛋白质，这些蛋白质的同时结合的影响 将在体外检查真菌。 表达模式，动力学 在表达和性质的决定因素的变化将被审查， 模式来发展表达调控的假说， 这种变化的机制。 抗体的能力 与表面组分反应将在研究中被利用 真菌对组织切片粘附中宿主-寄生虫相互作用 小鼠脾、肝、淋巴结、肾等。正常组织， 免疫功能低下和细胞毒性药物治疗的动物将 审查以评估东道国的状况对程度的影响， 绑定的本地化。 抗体和其他蛋白质结合到 表面将被用来选择突变体的表达改变， 表面表位和这些突变体的特征在于表面表达， 组织粘附。

DESCRIPTION (adapted from applicant's abstract): Candida albicans is an opportunistic dimorphic fungus capable of causing devastating disease in humans. Recent studies suggest that the surface topography of the cell is dynamic rather than static and that changes occur not just on cells grown under different conditions, but on single cells in the absence of cell division. Such dynamics reflect on mechanisms of synthesis and regulation of cell wall components and their assembly into a functional structure. They also impact on the interactions between the organism and host in health and disease since many of these interactions are mediated at the cell surface. This proposal focuses on the mannoproteins and proteins, which are the least well characterized constituents and have potential for great diversity. These components have been shown as a class to be present at the surface and implicated in the adherence of the fungus to host tissue and binding of host proteins. The research will take the approach of producing monoclonal antibodies to both carbohydrate and protein portions of these components to identify surface moieties. Antibodies will also be produced to constituents associated with binding of host proteins and fungal morphogenesis. The reactivity of these antibodies will be determined with whole cells using indirect immunofluorescence. The nature and identity of the antigenic determinants 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 and germ tubes will be determined. Since the organism is exposed to multiple antibodies and host proteins in the host, the affect of simultaneous binding of these to fungi will be examined in vitro. 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 interaction in adherence of fungi to tissue sections of murine spleen, liver, lymph node, kidney, etc. Tissues from normal, immunocompromised and cytotoxic antineoplastic drug treated animals will be examined to assess the effect of the state of the host on the extent and localization of binding. Antibodies and other proteins binding to the surface will be utilized to select for mutants altered in expression of surface epitopes and these mutants characterized for surface expression and tissue adherence.