Plague vaccine candidates by a functional genomic screen

描述（由申请人提供）：我们工作的长期目标是生产一套全面的疫苗，可以有效地保护大量人口免受基因工程病原体的侵害。这里描述的建议的直接目标是筛选鼠疫耶尔森氏菌的全基因组，以获得最有效的候选疫苗。鼠疫是一个重要的目标，至少有两个原因。这是一种极其致命的疾病，自古以来就被人类所感染。2)迫切需要生产一种广泛有效、安全、廉价的疫苗来应对生物恐怖主义的威胁。目前正在研究的亚单位疫苗的免疫原性简单，可能会限制其保护不同人群的能力，并可能使其对转基因病原体无效。我们第一阶段的具体目标是1)建立所需的信息学，以支持对整个鼠疫菌基因组的保护性抗原进行技术要求高的功能性筛选；2)确定生产完整鼠疫菌基因库的方案；3)通过在免疫和保护试验中检测鼠疫菌基因，确定最佳基因剂量和基因组筛选设计。我们的实验方法结合了我们的三种技术：基因免疫，表达文库筛选和线性基因表达元件。在第二阶段，我们将应用该系统在阵列基质中评估每个鼠疫杆菌基因作为PCR产物以保护小鼠免受感染。阳性评分基因将构成抗原选择组合，以保护广大人群免受自然和非自然鼠疫杆菌感染。除了确定未来疫苗的成分外，这些保护性抗原还将揭示免疫保护标记，从而促进这些疫苗以及其他潜在鼠疫疫苗的临床验证。

DESCRIPTION (provided by applicant): The long-term objective of our work is to produce a comprehensive set of vaccines that would effectively protect a large population against even genetically-engineered pathogens. The immediate objective of the proposal described here is to screen the whole genome of Yersinia pestis for the most potent vaccine candidates. Plague is a significant target for at least two reasons. 1) It is an extremely virulent, often lethal disease that has been suffered by humans since antiquity. 2) There is an immediate need to produce a broadly effective, safe, and inexpensive vaccine to counter the threat of bioterrorism. The immunogenic simplicity of a subunit vaccine currently under study may limit its ability to protect the diverse human population, and may render it ineffectual against a genetically-modified pathogen. Our phase I specific aims are to i) establish the informatics required to support a technically demanding, functional screen of the entire Y. pestis genome for protective antigens, ii) identify protocols for producing a complete Y. pestis gene library iii) define the optimal gene dose and design for this genome screen by testing Y. pestis genes in immune and protection assays. Our experimental approach combines three of our technologies: genetic immunization, expression library screening, and linear gene expression elements. In phase II we will apply this system to evaluate every Y. pestis gene as a PCR product in an arrayed matrix for protection of mice against infection. Positively scoring genes will constitute a portfolio of antigenic options to protect a broad population against natural and unnatural Y. pestis infections. In addition to identifying the components for a future vaccine, these protective antigens will reveal immune markers of protection, and thereby facilitate clinical validation of these as well as other potential plague vaccines.