IMMUNOREACTIVE MACROMOLECULES OF COCCIDIOIDES CELL TYPES

球孢子菌病是一种系统性真菌感染常见于西南 美国的 大多数感染是自限性的，但有些患者 发生播散性球孢子菌病或持续性疾病， 可能会危及生命 我们的研究重点是 C.免疫球蛋白 这 包括刺激T细胞以及抗体应答的抗原。 这种综合方法的逻辑是基于我们目前对 宿主对球虫感染的免疫反应。 球孢子菌病是一种T细胞介导的免疫系统被破坏的疾病。 显示在宿主防御中起关键作用，但对 T细胞反应性大分子的精确性质。 早期发现 原发性球孢子菌病由沉淀素抗体反应指示 梭免疫性抗原 然而，再一次， 血清学反应性大分子未知。 解决这些 问题，我们已经使用的方法来确定候选分子的C。 免疫炎，其通过以下组合刺激T细胞或抗体应答 生物化学和重组DNA技术。 我们将首先确定C。 通过使用刺激细胞介导免疫的免疫大分子 抗原特异性鼠T细胞系。 候选分子通过以下方法获得： 复杂混合物的分馏先前被证明是 在细胞免疫测定中具有免疫反应性。 第二种方法强调， 在拟议的研究中，从mRNA来源的cDNA表达文库， C.在lambda ZAP中构建 II用抗上述免疫反应性抗体进行筛选 混合物。 鉴定编码免疫反应性融合的cDNA克隆 蛋白质最终会导致基因的分离， 转化到合适的表达载体中，用于体外或体内产生 T细胞反应蛋白。 我们对早期梅毒血清学诊断的研究 球孢子虫感染集中在一个120-Kda的糖蛋白， 3-O-甲基化的杂甘露聚糖 后者结合特异性抗C。 免疫性肠炎患者IgM，显然是独特的碳水化合物部分，这是 真菌是系统性真菌病原体之一。 我们在这一部分的具体目标 该项目的目的是分离，纯化和表征免疫反应性 细胞外120-Kda糖蛋白的寡糖亚组分 用于原发性球孢子菌病的血清学诊断。 我们的总体目标 是为了鉴定C.引起体液性免疫反应 和细胞免疫反应，希望能提高诊断 试剂，并最终开发出一种针对球孢子菌病的疫苗。

Coccidioidomycosis is a systemic fungal infection common to southwestern United States. Most infections are self-limiting, but some patients develop disseminated coccidioidomycosis or persistent disease, either one of which can be life threatening. Our focus in the proposed research is the characterization of immunoreactive macromolecules of C. immitis. This includes antigens which stimulate T-cells as well as antibody response. The logic for this integrated approach is based on our current knowledge of the immunological response of the host to coccidioidal infections. Coccidioidomycosis is a disease in which T-cell mediated immunity has been shown to play a critical role in host defense, but little is known of the precise nature of T-cell reactive macromolecules. Early detection of primary coccidioidomycosis is indicated by a precipitin antibody response to C. immitis antigen. Once again, however, the precise nature of the serologically-reactive macromolecule(s) is unknown. To address these problems we have used the approach of identifying candidate molecules of C. immitis which stimulate T-cell or antibody response by a combination of biochemical and recombinant DNA techniques. We will initially identify C. immitis macromolecules which stimulate cell-mediated immunity by using antigen-specific murine T-cell lines. Candidate molecules are obtained by fractionation of complex mixtures previously demonstrated to be immunoreactive in cellular immunoassays. In a second approach, emphasized in the proposed research, CDNA expression libraries derived from MRNA of saprobic and parasitic phases of C. immitis and constructed in lambda ZAP II are screened with antibody raised against the above immunoreactive mixtures. Identification of CDNA clones which encode immunoreactive fusion proteins can ultimately lead to isolation of genes that can be introduced into appropriate expression vectors for in vitro or in vivo production of T-cell reactive proteins. Our research on serodiagnosis of early coccidioidal infection has focused on a 120-Kda glycoprotein that consists of a 3-O-methylated heteromannan. The latter, which binds specific anti-C. immitis patient IgM, is apparently unique to carbohydrate fractions of this fungus among the systemic fungal pathogens. Our specific aims in this part of the project are to isolate, purify, and characterize the immunoreactive oligosaccharide subfractions(s) of the extracellular, 120-Kda glycoprotein for use in serodiagnosis of primary coccidioidomycosis. Our overall goals are to characterize specific antigens of C. immitis which elicit humoral and cellular immune responses in the hopes of improving diagnostic reagents, and eventually developing a vaccine against coccidioidomycosis.