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: 7675204
• 负责人: JOHN N GALGIANI
• 金额: $ 32.76万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7675204
• 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.

球虫剂属。是真菌病原体，通常会引起与大量相关的肺炎 发病率和随之而来的经济或医疗费用。某些感染会导致呼吸衰竭，柔软 组织脓肿，骨髓炎或脑膜炎。即使在健康的人中，一个孢子也吸入 足以导致其中一个或多个并发症死亡，这种感染程度是 在某种程度上负责美国和苏联的过去开发计划使用球虫剂 spp。作为生物武器。球球菌属属最近被NIAID归类为C类代理，也是 考虑了新兴的公共卫生病原体。 由于球虫性感染经常会产生终身保护免受再感染的保护，因此很可能 预防性疫苗可能是有效的。大约二十粒球菌的筛选程序 蛋白质鉴定出建议用于临床试验的重组疫苗。但是，制造业 与抗原的特定顺序相关的困难阻碍了其进一步的发展。在这个项目中，我们 应使用体外蛋白表达来允许大规模抗原筛查作为识别的手段 同样可以配方的保护性抗原。球形细胞的蛋白质组学分析 墙已经确定了650种蛋白质，其中我们将筛选约1,000个外显子，最少 与哺乳动物蛋白的相似性，使用第一静脉反应性和随后的淋巴细胞反应性 先前感染了球虫剂或用保护性全细胞疫苗免疫的小鼠。我们的目标是 确定可以开发用于临床测试的第二代重组疫苗候选者。 该项目还将使用串联质谱法开发抗原检测分析作为更多 早期山谷热的敏感诊断。我们最近在被感染小鼠的肺中检测到了数十只 球蛋白蛋白。其中三个与哺乳动物蛋白没有相似性，与其他相似性<45％ 真菌蛋白。我们将通过重组方法表达这些生物签名靶标，并使用纯化 蛋白质产生高亲和力抗体。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，以作为理性和的独立分析 直接荧光染色或ELISA方法的最佳设计可检测临床标本中的抗原。