Coccidioides Proteins as Vaccine Antigens and Diagnostic Biosignatures

球孢子菌属是真菌病原体，通常会引起与相当大的 发病率和随之而来的经济或医疗费用。有些感染会导致呼吸衰竭， 组织水肿、骨髓炎或脑膜炎。即使在其他方面健康的人，吸入单个孢子 足以导致这些并发症中的一种或多种死亡，这种程度的传染性 在某种程度上，美国和苏联过去的开发计划是使用球孢子菌 spp.作为生物武器。球孢子菌属，最近被NIAID列为C类代理，也是 被认为是新兴的公共卫生病原体。 由于球孢子虫感染经常产生终身保护，防止再感染，很可能是一个 预防性疫苗是有效的。一个筛选计划，大约24个球孢子菌 蛋白质鉴定了一种重组疫苗，推荐用于临床试验。然而，制造业 与抗原特异性序列相关的困难阻碍了其进一步发展。本课题 应使用体外蛋白表达以允许更大规模的抗原筛选作为鉴定 同样具有保护性的抗原更易于配制。小球细胞的蛋白质组学分析 Walls已经鉴定了650种蛋白质，我们将筛选其中大约1,000个外显子， 与哺乳动物蛋白质的相似性，首先使用来自 先前用球孢子菌感染或用保护性全细胞疫苗免疫的小鼠。我们的目标是 鉴定可开发用于临床试验的第二代重组疫苗候选物。 该项目还将使用串联质谱法来开发抗原检测测定法， 早期山谷热的敏感诊断我们最近在受感染的小鼠的肺部检测到数十种 球孢子菌蛋白其中三个与哺乳动物蛋白质没有相似性，与其他蛋白质的相似性<45%。 真菌蛋白我们将通过重组方法表达这些生物特征靶标，并使用纯化的 蛋白质来产生高亲和力抗体。串联质谱法也将用于表征 抗原-抗体相互作用的强度和特异性，作为合理和 优化设计直接荧光染色或ELISA方法检测临床标本中的抗原。

Coccidioides spp. are fungal pathogens that normally causes a pneumonia associated with considerable morbidity and attendant economic or medical care costs. Some infections produce respiratory failure, soft tissue abscesses, osteomyelitis or meningitis. Even in otherwise healthy persons, inhalation of a single spore is sufficient to result in death from one or more of these complications, and this degree of infectivity was responsible, in part, for past development programs by the U.S. and the Soviet Union to use Coccidioides spp. as biological weapons. Coccidioides spp., recently classified as Category C agents by NIAID, are also considered emerging public health pathogens. Because coccidioidal infection so often produces life-long protection against reinfection, it is likely that a preventative vaccine could be effective. A screening program of approximately two dozen coccidioidal proteins identified a recombinant vaccine that was recommended for clinical trials. However, manufacturing difficulties related to the antigen's specific sequence has impeded its further development. In this project, we shall use in vitro protein expression to permit antigen screening on a larger scale as a means of identifying equally protective antigens that are more amenable to formulation. A proteomic analysis of spherule cell walls has identified 650 proteins and of these we shall screen approximately 1,000 exons with the least similarity to mammalian proteins, using first seroreactivity and subsequently reactivity of lymphocytes from mice previously infected with Coccidioides or immunized with protective whole cell vaccines. Our goal is to identify a second-generation recombinant vaccine candidate that could be developed for clinical testing. This project will also use tandem mass spectrometry to develop antigen-detecting assays as a more sensitive diagnostic for early Valley Fever. We have recently detected in the lungs of infected mice dozens of coccidioidal proteins. Three of these have no similarity to mammalian proteins and <45% similarity to other fungal proteins. We shall express these biosignature targets by recombinant methods and use the purified proteins to raise high-affinity antibodies. Tandem mass spectrometry will also be used to characterize the strength and specificity of antigen-antibody interactions to serve as independent analyses for the rational and optimal design of direct fluorescent staining or ELISA methods to detect antigens in clinical specimens.