Host control in Coccidioidomycosis

描述(申请人提供)：球孢子菌病是一种全国性的公共卫生问题，在亚利桑那州具有特殊的严重性，对于患有艾滋病或其他免疫抑制的患者、怀孕期间的妇女和一些少数民族来说。它给军方和球孢子虫带来了问题。是对国土安全的潜在威胁。目前的治疗方法是不够的。亚利桑那大学拥有位于流行区域内的唯一一所医学院，能够支持生物科学的研究，并持续致力于改善我们对球孢子菌病的控制。这种MRU方案的统一方法是在研究感染机体和受感染宿主的适应性反应的同时，识别和验证球孢子虫的靶抗原。项目1将提高我们评估可能进行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，使得它们的解释含糊不清。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性的组织学模式。我们还将研究人类和小鼠的组织反应之间的相似性。项目2将侧重于真菌本身如何参与宿主的疾病恢复。我们假设，静止的球状状态是由特殊的基因表达产生的，类似于分生孢子形成后的静止或其他休眠的腊肠结构。基因的表达将通过分析体外培养的球藻的长SAGE文库来确定。扩展到对敏感和耐药小鼠的体内研究。通过这些方法选择的与静止相关的真菌基因将通过根癌农杆菌介导性转化而被破坏。负责静默的基因表达可以识别新的靶点，以利于治疗。项目3打算为广泛传播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在体外可以用树突状细胞(DC)逆转。为了将这一进展转化为实际治疗，需要通过已定义的重组抗原产生类似的效果。该项目将使用DC作为载体来评估使用CPGS和MPL等佐剂递送重组抗原的条件。我们还将分析对抗原冲击的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.