Host control in Coccidioidomycosis

描述（由申请人提供）：球孢子菌病是亚利桑那州特别严重的全国性公共卫生问题，主要针对艾滋病患者或其他免疫抑制患者、孕期妇女和一些少数民族。它给军队带来了问题，而且球虫是对国土安全的潜在威胁。目前的治疗方法是不够的。亚利桑那大学拥有唯一一所位于流行地区的医学院，可以支持生物科学研究，并一直致力于改善我们对球孢子菌病的控制。该MRU提案的统一方法是识别和验证球虫靶抗原，同时研究感染生物体和被感染宿主的适应性反应。项目1将提高我们评估可能用于临床试验的候选疫苗的能力。目前的评估高度依赖于实验条件，使其解释模糊不清。项目1将使用免疫组织化学、蛋白质组学分析和原位杂交来确定遗传和获得性耐药的组织学模式。我们还将研究人类和小鼠的组织学反应之间的相似性。项目2将重点关注真菌本身如何参与宿主从疾病中恢复。我们假设静止的球粒状态是由特殊的基因表达产生的，类似于分生后的静止或其他休眠的腐殖结构。基因表达将通过分析体外培养球虫的长sage文库来确定，并扩展到体内易感和耐药小鼠的研究。通过这些方法选择的与静止相关的真菌基因将被农杆菌介导的转化破坏。负责静息的基因表达可以确定新的治疗靶点。项目3旨在开发一种广泛传播的球孢子菌病的免疫治疗方法。在体外树突状细胞（DC）可以逆转球孢子菌病的抗原特异性能量。为了将这一进展转化为实际治疗，需要通过确定重组抗原来引发类似的效果。该项目将评估使用DC作为载体递送带有CpGs和MPL等佐剂的重组抗原的条件。我们还将分析对抗原脉冲DC有反应的淋巴细胞亚群的表型、功能和细胞因子谱。

DESCRIPTION (provided by applicant): Coccidioidomycosis is a national public health problem with special intensity to Arizona, for patients with AIDS or other immunosuppression, women during pregnancy, and some ethnic minorities. It poses problems for the military and Coccidioides spp. are a potential threat to homeland security. Current therapies are inadequate. The University of Arizona has the only medical school situated within the endemic region, can support research in the biological sciences, and has an ongoing commitment improve our control of coccidioidomycosis. The unifying approach of this MRU proposal is to identify and validate Coccidioides target antigens while studying the adaptive responses of both the infecting organism and the infected host. Project 1 will improve our ability to evaluate vaccine candidates for possible clinical trials. Current assessments are highly dependent upon experimental conditions, making their interpretation ambiguous. Project 1 will use immunohistochemistry, proteomic analyses, and in situ hybridization to determine the histologic patterns of genetic and acquired resistance. We shall also examine similarities between the histological response in humans and mice. Project 2 will focus on how the fungus itself participates in the host's recovery from illness. We hypothesize that a quiescent spherule state is produced by specialized gene expression analogous to the quiescence after conidiation or other dormant saprobic structures. Gene expression will be determined from analysis of long-SAGE libraries of in vitro grown Coccidioides spp. with extension to in vivo studies of susceptible and resistant mice. Selected fungal genes associated with quiescence by these methods will be disrupted using Agrobacterioum tumefaciens-mediated transformation. Gene expression responsible for quiescence could identify novel targets for therapeutic benefit. Project 3 intends to develop an immunologic therapy for patients with widely disseminated coccidioidomycosis. Antigen-specific anergy in coccidioidomycosis can be reversed in vitro with dendritic cells (DC). To translate this advance to practical therapy, similar effects will need to be elicited by defined recombinant antigens. This project will evaluate conditions using DC as a vehicle to deliver recombinant antigens with adjuvants such as CpGs and MPL. We will also analyze the phenotype, function and cytokine profiles of lymphocyte subsets that respond to antigen-pulsed DC.