Coccidioides Proteins as Vaccine Antigens and Diagnostic Biosignatures

球孢子菌蛋白作为疫苗抗原和诊断生物特征

• 批准号: 8260265
• 负责人: JOHN N GALGIANI
• 金额: $ 34万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8260265
• 关键词: Abscess; Amino Acid Sequence; Animals; Antibodies; Antibody Affinity; Antigens; Biological; Biological Assay; Blood Tests; Breathing; Categories; Cell Wall; Cessation of life; Chimeric Proteins; Clinical; Clinical Trials; Coccidioides; Coccidioidomycosis; Collaborations; Cysteine; Data; Development; Diagnostic; Diagnostic tests; Disease; Drug Formulations; Economics; Effectiveness; Emerging Communicable Diseases; Enzyme-Linked Immunosorbent Assay; Exons; Fever; Fungal Antigens; Fungal Proteins; Future; General Population; Generations; Genes; Goals; Immunoassay; Immunologics; In Vitro; Infection; Infectious Diseases Research; Injection of therapeutic agent; Life; Lung; Lymphocyte; Macaca fascicularis; Medical Care Costs; Meningitis; Methods; Morbidity - disease rate; Mus; National Institute of Allergy and Infectious Disease; Osteomyelitis; Persons; Phase; Pneumonia; Production; Program Development; Protein Microchips; Proteins; Proteomics; Public Health; Quality Control; Recombinant Proteins; Recombinant Vaccines; Recombinants; Reproduction spores; Resistance; Resolution; Respiratory Failure; Saccharomyces cerevisiae; Screening procedure; Serologic tests; Site; Specificity; Specimen; Staining method; Stains; Testing; Time; Tissues; USSR; Uncertainty; Vaccination; Vaccine Antigen; Vaccines; Whole Cell Vaccine; base; biodefense; biosignature; clinical Diagnosis; design; disulfide bond; editorial; immunoreactivity; migration; pathogen; prevent; programs; protein aggregation; protein expression; research clinical testing; soft tissue; tandem mass spectrometry; vaccine candidate; vaccine development; weapons

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.

球孢子菌属是真菌病原体，通常会引起与相当大的相关的肺炎 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、软 组织脓肿、骨髓炎或脑膜炎。即使在其他方面健康的人中，吸入单个孢子 足以导致因这些并发症中的一种或多种而死亡，并且这种程度的传染性 部分负责美国和苏联过去使用球孢子菌的开发计划 种。作为生物武器。 Coccidioides spp. 最近被 NIAID 分类为 C 类药物，也属于 被认为是新出现的公共卫生病原体。 由于球孢子菌感染常常能产生终生的保护，防止再次感染，因此很可能 预防性疫苗可能有效。大约两打球孢子菌的筛查计划 蛋白质鉴定出一种重组疫苗，建议进行临床试验。然而，制造 与抗原的特定序列相关的困难阻碍了其进一步发展。在这个项目中，我们 应使用体外蛋白质表达来进行更大规模的抗原筛选，作为识别的手段 具有同等保护性的抗原，更易于配制。球细胞的蛋白质组学分析 walls 已鉴定出 650 种蛋白质，其中我们将筛选大约 1,000 个具有最少的外显子 与哺乳动物蛋白质的相似性，首先使用血清反应性，然后使用来自淋巴细胞的反应性 先前感染球孢子菌或用保护性全细胞疫苗免疫的小鼠。我们的目标是 确定可用于临床测试的第二代重组候选疫苗。 该项目还将使用串联质谱法来开发抗原检测方法，作为更有效的方法。 早期谷热的敏感诊断。我们最近在受感染小鼠的肺部发现了数十个 球孢子蛋白。其中三种与哺乳动物蛋白质没有相似性，与其他蛋白质的相似性<45% 真菌蛋白质。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 蛋白质以产生高亲和力抗体。串联质谱也将用于表征 抗原-抗体相互作用的强度和特异性，可作为合理和合理的独立分析 优化设计直接荧光染色或 ELISA 方法来检测临床标本中的抗原。