Early in vivo expressed antigens and their role in virulence, immune response, and vaccines for coccidioidomycosis

项目摘要/摘要(动物核心) 免疫球虫和波斯达氏球虫引起球孢子菌病(谷热-VF)，这是非常令人关注的 由于最近美国疾病发病率迅速上升。我们计划(VIRV-CCRC)的总体目标是- 研究在VF感染周期早期高表达的蛋白质，当真菌节孢子是 发育成球状，形成内孢子。我们在研究项目1(RP1)中的假设是PRO- 在这一关键阶段表达的teins是疾病进展的关键，并且可能代表毒力因子。 动物核心(ANC)将通过提供新的动物模型来解决这一假说 毒力测试。在目标1中，我们已经展示的Galeria(蜡虫)无脊椎动物模型支持节肢动物- 分生孢子发育成成熟的球状孢子代表了对这一步至关重要的致病因子的良好测试。它是 同样经济，并将减少研究所需的小鼠数量。尽管如此，鼠标模型PRO- 提供Galleria不能提供的数据，非国大将提供这些数据来确认Galleria的毒力数据。在Aim 2中，鼠标 挑战实验将在北亚利桑那大学的ABSL3设施中进行。身份识别 将使用早期感染毒力因子来指导T细胞受体分析中T细胞表位的鉴定。 RP2中的SIS。球虫抗原表位的T细胞识别是免疫应答中的关键步骤，我们将对此进行研究。 被擦除成新的VF诊断测试。毒力因子和T细胞受体结果都将指导优先考虑 推动先进的核酸(RNA和DNA)疫苗的开发。RP3将使用鼠标模型和非 人类灵长类动物模型来开发和测试新的疫苗。在目标3中，我们将培育一种新的猪尾猕猴 (NHP)模型。我们对这一模型的关注是由于他们已建立的室颤易感性和天然药物的可用性。 从亚利桑那州繁殖群体(WaNPRC)召集受感染的群体动物。AZ猕猴疾病发病率 与亚利桑那州人群相似，代表了病理数据的宝贵来源，血清，PBMC， 和其他纸巾。富勒实验室(RP3)已经成功地将猪尾猕猴用于疫苗接种- IES，实现了高抗体效价和对其他疾病的保护性免疫。非国大将协调VAC- 电影工作通过亚利桑那州自然感染的NHP人群，西雅图WaNPRC设施为NHP IM- 市民化研究，以及杜兰国家灵长类研究中心对球虫的挑战研究 NHP。总而言之，非国大将开发、提供和支持三种互补的动物模型：Galleria、 鼠标和NHP。每一项都有其独特的优势，支持了 VIRV-CCRC。非国大是一项跨越三个机构和四个地点的协调努力，利用 国家优势，以最大限度地提高整个VIRV-CCRC的生产力。随着VIRV-CCRC培育出这些动物 模型资源，我们将鼓励与其他CCRC合作，并广泛地与山谷发烧研究合作 社区。高质量的动物模型和组织标本将缓解一些研究障碍 到开发更好的治疗剂、诊断测试和预防性疫苗。

Project Summary/Abstract (Animal Core) Coccidioides immitis and C. posadasii cause coccidioidomycosis (Valley Fever - VF), which is of great concern due to recent rapid increases in US disease incidence. The overall goal of our program (VIRV-CCRC) is to in- vestigate proteins that are highly expressed early in the VF infection cycle, when the fungal arthroconidia are developing into spherules and endospores forming. Our hypothesis in Research Project 1 (RP1) is that pro- teins expressed at this critical stage are key to disease progression and potential represent virulence factors. The Animal Core (AnC) will be instrumental in addressing this hypothesis by providing new animal models for virulence testing. In Aim 1, the Galleria (wax worm) invertebrate model that we have shown supports arthro- conidia development into mature spherules represents a good test of virulence factors key to this step. It is also economical and will reduce the number of mice needed for research. Nevertheless, the mouse model pro- vides data that Galleria cannot and the AnC will provide it to confirm Galleria virulence data. In aim 2, mouse challenge experiments will be conducted at the ABSL3 facility at Northern Arizona University. Identification of early infection virulence factors will be used guide the identification of T cell epitopes in a T cell receptor analy- sis in RP2. T cell recognition of Coccidioides epitopes is a key step in the immune response, which we will lev- eraged into novel VF diagnostic tests. Both virulence factor and T cell receptor results will guide the prioritiza- tion of advanced nucleic acid (RNA and DNA) vaccine development. RP3 will use the mouse model and a non- human primate model to develop and test new vaccines. In aim 3, we will develop a new pig-tailed macaque (NHP) model. Our focus on this model is due to their established VF susceptibility and the availability of natu- rally infected colony animals from the Arizona Breed Colony (WaNPRC). The AZ macaque disease incidence is similar to the Arizona human population and represents a valuable source of pathology data, serum, PBMC, and other tissues. The Fuller lab (RP3) has already successfully used pig-tailed macaques in vaccination stud- ies, achieving high antibody titers and protective immunity to other diseases. The AnC will coordinate the vac- cine work through the naturally infected NHP population in Arizona, the Seattle WaNPRC facility for NHP im- munization studies, and at Tulane National Primate Research Center for Coccidioides challenge studies in NHP. In summary, the AnC will develop, provide, and support three complementary animal models: Galleria, mouse, and NHP. Each has its distinct advantages that support the individual and overall research goals of the VIRV-CCRC. The AnC is a coordinated effort across three institutions and four locations that leverages institu- tional strengths to maximize productive of the whole VIRV-CCRC. As the VIRV-CCRC develops these animal model resources, we will encourage collaboration with other CCRC and broadly with the Valley Fever research community. High quality animal models and their tissue specimens will alleviate some of the research barriers to developing better therapeutic agents, diagnostic tests, and preventive vaccines.