Coccidioides Proteins as Vaccine Antigens and Diagnostic Biosignatures

球藻(Coccidioidesspp.)通常引起肺炎的真菌病原体是否与 发病率和随之而来的经济或医疗费用。一些感染会导致呼吸衰竭，软 组织脓肿、骨髓炎或脑膜炎。即使在其他健康的人中，吸入单一的孢子 足以导致这些并发症中的一种或多种死亡，而这种传染性程度是 对美国和苏联过去使用球菌素的发展计划负有部分责任 SPP.作为生物武器。球藻属(Coccidioidesspp.)，最近被NIAID归类为C类细菌，也是 被认为是新出现的公共卫生病原体。 由于球虫感染经常产生终身保护，防止再次感染，很可能是一种 预防性疫苗可能是有效的。大约20多个球虫的筛查计划 Proteins确定了一种推荐用于临床试验的重组疫苗。然而，制造业 与抗原特定序列相关的困难阻碍了它的进一步发展。在这个项目中，我们 应使用体外蛋白表达来进行更大规模的抗原筛选，作为鉴定的一种手段 同样具有保护性的更易于配制的抗原。球状细胞的蛋白质组学分析 WALS已经鉴定了650种蛋白质，我们将用最少的方法筛选大约1000个外显子 与哺乳动物蛋白质的相似性，使用第一次血清反应性和随后的淋巴细胞反应性 以前感染过球虫或用保护性全细胞疫苗免疫的小鼠。我们的目标是 确定可开发用于临床测试的第二代重组疫苗候选疫苗。 该项目还将使用串联质谱仪来开发抗原检测分析，作为一种 对早期谷热的敏感诊断。我们最近在受感染的小鼠的肺中检测到数十种 球虫蛋白。其中三个与哺乳动物的蛋白质没有相似性，而与其他蛋白质的相似性为45% 真菌蛋白质。我们将通过重组的方法表达这些生物标志物，并使用纯化的 产生高亲和力抗体的蛋白质。串联质谱学也将被用于表征 抗原-抗体相互作用的强度和特异性可作为对理性和 直接荧光染色或酶联免疫吸附试验检测临床标本中抗原的优化设计

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.