CELL SURFACE DYNAMICS OF MANNOPROTEINS OF C ALBICANS

简介(摘自申请者摘要)：白色念珠菌是一种 一种能引起毁灭性疾病的机会性二态真菌 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，变化不仅仅发生在培养的细胞上 在不同的条件下，但在单个细胞上没有细胞 组织。这种动态反映了合成和调节的机制。 细胞壁组件及其组装成功能结构。 它们还会影响生物体和寄主之间的相互作用 健康和疾病，因为这些相互作用中的许多是在 细胞表面。这项建议关注的是甘露糖蛋白和蛋白质， 它们是特征最差的成分，有可能 多样性很强。这些组件已显示为要呈现的类 在表面，与真菌与宿主组织的粘连有关 和宿主蛋白的结合。 这项研究将采取生产单抗的方法来 对这些成分的碳水化合物和蛋白质部分进行鉴定 表面部分。抗体也会对成分产生。 与宿主蛋白的结合和真菌的形态发生有关。这个 这些抗体的反应性将在整个细胞中使用 间接免疫荧光。抗原性和同一性 将使用免疫印迹技术检查决定因素，敏感性 各种试剂和其他技术的抗原。其影响范围 生长和静止相酵母菌和芽管的表位表达 将会被确定。因为有机体暴露在多种抗体下 和寄主蛋白，它们同时结合的影响 对真菌的影响将在体外进行检测。表达的模式，动力学 将检查行列式的表达和性质的变化 发展表达调控假说的模式 实现这类变化的机制。抗体的能力 将在研究中利用与表面组分的反应 真菌黏附组织切片中的寄主-寄生虫相互作用 小鼠脾、肝、淋巴结、肾等。 免疫受损和细胞毒性抗肿瘤药物治疗的动物将 检查以评估主机状态对范围的影响和 绑定的本地化。抗体和其他结合在一起的蛋白质 将利用表面来筛选在表达中改变的突变体 表面表位和这些突变体的表面表达和 组织粘连。

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.