IMMUNOREACTIVE MACROMOLECULES OF COCCIDIOIDES CELL TYPES

粗球孢子菌是人类的真菌呼吸道病原体， 山谷热（球孢子菌病）的病原体。暴露于 空气传播的感染性细胞（节孢子）可导致一系列 临床疾病由一种良性、自限性的感染性细胞引起 （节分生孢子）可导致一系列临床疾病， 一种严重的、进行性的、通常致命的真菌病的自限性感染 包括肺和肺外组织。免疫 危害性疾病，虽然存在于一些患者中， 感染C.免疫球蛋白存在一个一致的， 可预测的免疫模式，患者在不同阶段的 球孢子菌病，这可能与初始暴露于 关节分生孢子，然后反复暴露于可溶性抗原 寄生循环期间主要壁起源的部分（即， 小球-内生孢子阶段）。我们的假设是节孢子， 小球和内生孢子。免疫反应引起不同的宿主反应 由于抗原性的定性和/或定量差异， 它们的细胞壁的组成，宿主最初暴露于其中。 已经有可能从细胞中分离可溶性抗原复合物， 体外生长的节分生孢子和小球的细胞被膜， 在体液和细胞免疫测定中表征它们的免疫反应性， 并使用各种分离技术获得纯化的组分。在 在拟议的调查中，我们将描述这些纯化的 成分的基础上，其化学成分，抗原性 标准化免疫电泳和免疫扩散试验， 在测定患者抗体的试验中的相对免疫反应性 吸附和T细胞对均质级分的反应。我们还将 研究这些纯化成分的生物学功能，重点是 它们可能的酶活性和对真菌形态发生的影响 和宿主入侵我们最近开发的cDNA表达文库 以及C.移民许可证 重组DNA技术在我们进一步表征中的应用 这些纯化的，与壁相连的部分。首先，我们将筛选 使用选择的免疫探针和寡核苷酸探针的文库 对应于具有确定的显著性的那些选定的级分 在宿主抗体应答、组织侵袭或C.免疫组织形态发生， 并克隆调控同源融合产物的基因 缩氨酸最终，我们将确定这些基因的表达， 节孢子-小球-内生孢子体外发育。我们认为 这种全面的方法来研究C.免疫抗原会导致 为了更好地了解免疫反应的模式， 宿主和致病因素，并确定 潜在的诊断DNA探针和分子靶点， 新的抗真菌化合物。

Coccidioides immitis is a fungal respiratory pathogen of humans and causative agent of valley fever (coccidioidomycosis). Exposure to the airborne, infectious cells (arthroconidia) can lead to a range of clinical disease from a benign, self-limited infectious cells (arthroconidia) can lead to a range of clinical disease from a benign, self-limited infection to a severe, progressive and often fatal mycosis involving pulmonary and extrapulmonary tissues. Immunologically compromising illnesses, although present in some patients, are not prerequisite to infection by C. immitis. There exists a consistent and predictable immunological pattern in patients at various stages of coccidioidomycosis, which may be related to initial exposure to the arthroconidia, followed by repeated exposure to soluble antigenic fractions of primarily wall origin during the parasitic cycle (i.e., spherule-endospore phase). Our hypothesis is that arthroconidia, spherules, and endospores of C. immitis elicit different host responses because of qualitative and/or quantitative differences in the antigenic composition of their cell wall to which the host is originally exposed. It has been possible to isolate soluble, antigenic complexes from the cell envelopes off arthroconidia and spherules grown in vitro, characterize their immunoreactivity in humoral and cellular immunoassays, and obtain purified components using various separation techniques. In the proposed investigations we will characterize these purified components on the basis of their chemical composition, antigenic identity in standardized immunoelectrophoresis and immunodiffusion tests, and relative immunoreactivity in assays which measure patient antibody adsorption and T-cell response to the homogeneous fractions. We will also examine the biological function of these purified components, with focus on their possible enzymatic activity and impact on fungal morphogenesis and host invasion. Our recent development of a cDNA expression library and the availability of a genomic library for C. immitis permits application of recombinant DNA technology in our further characterization of these purified, wall-associated fractions. Initially, we will screen the libraries using selected immunoprobes and oligonucleotide probes corresponding to those selected fractions with established significance in host antibody response, tissue invasion, or C. immitis morphogenesis, and clone the genes which regulate production off the homologous fusion peptides. Ultimately, we will determine expression of these genes during arthroconidium-spherule-endospore development in vitro. We believe that this comprehensive approach to the study of C. immitis antigens will lead to a better understanding of the pattern of immunological response by the host and factors which contribute to pathogenicity, as well as identify potential diagnostic DNA probes and molecular targets for development of novel antifungal compounds.