PATHOGENESIS OF ACANTHAMOEBA KERATITIS

我们将试图阐明发展的机制， 阿米巴角膜炎的细胞膜分子检测 角膜上皮和阿米巴。 在项目1中，我们将（1）确定 并表征这些兔角膜上皮细胞表面 在宿主-寄生虫相互作用中发挥作用的糖缀合物，（2） 确定寄生虫在阿米巴反应细胞上的附着部位 兔角膜上皮表面糖缀合物，（3）制备 针对这些阿米巴的单克隆和多克隆抗体， 角膜上皮细胞表面糖缀合物，其在角膜上皮细胞的生长中起作用。 宿主-寄生虫相互作用，（4）用免疫组织化学测定 技术，无论是阿米巴反应性糖缀合物，发现对兔 培养的角膜上皮细胞存在于正常人身上， 兔角膜在体内，和（5）确定是否有可能阻断 使用上述方法， 抗体和各种酶、外切糖苷酶和凝集素。 由于神经氨酸酶的许多病原体，包括流感病毒和 寄生虫克氏锥虫，在 感染，我们将在项目2中尝试确定阿米巴是否 神经氨酸酶在阿米巴角膜炎的发病机制中起作用。 在这项研究中，我们将（1）分离和表征神经氨酸酶 由人角膜分离的阿米巴原虫产生，并确定是否 该酶在结构上与其他神经氨酸酶相关， 寄生虫，特别是克氏锥虫，（2）识别那些兔角膜 上皮细胞表面糖缀合物，其作为 阿米巴神经氨酸酶和确定是否唾液酸化的角膜 上皮细胞表面糖复合物被寄生虫酶暴露 寄生虫附着位点和（3）确定是否 抗神经氨酸酶抗体和神经氨酸酶的非免疫抑制剂 抑制或增强阿米巴对人和兔的粘附 角膜上皮，从而确定阿米巴是否 神经氨酸酶，像克氏锥虫酶一样， 负控制机制 希望拟议的研究将导致更好地了解 阿米巴角膜炎的分子基础， 更有效的治疗方法来治疗这种威胁和困难的视力， 治疗疾病。

We shall attempt to elucidate the mechanism of the development of Acanthamoeba keratitis by examining the plasma membrane molecules of corneal epithelium and Acanthamoeba. In project 1 we shall (1) identify and characterize those rabbit corneal epithelial cell surface glycoconjugates which play a role in the host-parasite interactions, (2) pinpoint the sites of parasite attachment on Acanthamoeba-reactive cell surface glycoconjugates of rabbit corneal epithelium, (3) prepare monoclonal and polyclonal antibodies against those Acanthamoeba and corneal epithelial cell surface glycoconjugates which play a role in the host-parasite interactions, (4) determine using immunohistochemical techniques, whether Acanthamoeba-reactive glycoconjugate, found on rabbit corneal epithelial cells in culture are present on normal human and rabbit corneas in vivo, and (5) determine whether it is possible to block the adherence of Acanthamoeba to human and rabbit corneas using the above antibodies and by various saccharides, exoglycosidases and lectins. Since the neuraminidase of many pathogens, including influenza virus and parasite Trypanosoma cruzi, plays a role in the pathogenesis of infection, we shall, in project 2, try to determine whether Acanthamoeba neuraminidase plays a role in the pathogenesis of Acanthamoeba keratitis. In this study, we shall (1) isolate and characterize the neuraminidase produced by human corneal isolates of Acanthamoeba and establish whether the enzyme is structurally related to the neuraminidases of other parasites, especially T cruzi, (2) identify those rabbit corneal epithelial cell surface glycoconjugates which serve as substrates for Acanthamoeba neuraminidase and determine whether asialation of corneal epithelial cell surface glycoconjugates by the parasite enzyme exposes the parasite attachment sites and (3) determine whether anti-neuraminidase antibodies and non-immune inhibitors of neuraminidases inhibit or enhance the adherence of Acanthamoeba to human and rabbit corneal epithelium and thereby establish whether Acanthamoeba neuraminidase, like the T cruzi enzyme, regulates the infection by a negative control mechanism. It is hoped that the proposed studies will lead to a better understanding of the molecular basis of Acanthamoeba keratitis and to the development of more effective therapy for this sight threatening and difficult to treat disease.