Host control in Coccidioidomycosis

描述（由申请人提供）：球孢子菌病是一种全国性的公共卫生问题，对亚利桑那州的艾滋病或其他免疫抑制患者、怀孕期间的妇女和一些少数民族具有特殊的强度。它给军队和球孢子菌带来了问题。是对国土安全的潜在威胁目前的治疗是不够的。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，并不断致力于改善我们对球孢子菌病的控制。该MRU建议的统一方法是在研究感染生物体和受感染宿主的适应性反应的同时，识别和验证球孢子菌靶抗原。项目1将提高我们为可能的临床试验评估候选疫苗的能力。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的疾病康复。我们推测，一个静止的小球状态是由专门的基因表达类似于孢子形成或其他休眠saprobic结构后的静止。基因表达将通过分析体外生长的球孢子菌属物种的长SAGE文库来确定。并扩展到易感小鼠和抗性小鼠的体内研究。将使用根癌农杆菌介导的转化来破坏通过这些方法选择的与休眠相关的真菌基因。负责静止的基因表达可以确定新的治疗目标。项目3旨在为广泛播散的球孢子菌病患者开发一种免疫疗法。球孢子菌病的抗原特异性无能可以在体外用树突状细胞（DC）逆转。为了将这一进展转化为实际治疗，需要通过确定的重组抗原引起类似的效果。该项目将评估使用DC作为载体递送具有佐剂如CpG和MPL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.