Identification of New Antigens for a Plague Vaccine

描述（由申请人提供）：鼠疫耶尔森氏菌是急性疾病腺鼠疫和肺鼠疫的病原体，也是人类经历的最具破坏性的致病细菌之一，现在被世界卫生组织归类为重新出现的人类病原体。传染的可能性，缺乏有效的疫苗，以及多种抗生素耐药菌株的出现，使Y。在美国，鼠疫是一种潜在的生物恐怖主义制剂。我们的长期目标是阐明Y.鼠疫更直接的目标是鉴定和评估新的和现有的Y抗原。研制新一代鼠疫疫苗。Y.鼠疫是已知的。提出了四个目标。在目的1中，我们将制备Y的Braun/murein脂蛋白（lpp）-负突变体。鼠疫，基于我们最近的数据，即鼠伤寒沙门氏菌和假结核耶尔森氏菌的专利Ipp同基因突变体在小鼠中无毒力，并提供保护，免受野生型细菌的攻击。我们将在小鼠中检测这些突变体的免疫应答，以开发活的减毒Y。鼠疫疫苗或使用Y.假结核病是Y.鼠疫抗原目的二是鉴定Y染色体上差异表达或特异表达的基因（可能与毒力相关）。鼠疫的基因组学和蛋白质组学，以评估用于重组亚单位鼠疫疫苗的新抗原。目的3研究筛选出的在体内表达的基因的毒力潜力。通过开发等基因突变体并评估它们在小鼠模型中的致死性，评估所选抗原提供针对鼠疫杆菌的免疫力的能力，鼠疫挑战目的4将研究Y的Ipp负突变体的潜在用途。pseudotuberculosis和S.鼠伤寒沙门氏菌作为载体传递Y.鼠疫抗原，通过从诱导型启动子下的质粒和/或减毒Y.假结核/S.鼠伤寒沙门氏菌或DMA疫苗接种。或者，S. Typhi（Ty 21 a）也可以使用。我们相信，这些多种方法将确定候选抗原用于一个新的，有效的鼠疫疫苗。

DESCRIPTION (provided by applicant): Yersinia pestis, an etiological agent of the acute diseases bubonic and pneumonic plague and one of the most devastating epidemic-causing bacteria experienced by mankind, is now classified as a re-emerging human pathogen by the WHO. The potential for contagion, lack of an effective vaccine, and emergence of multiple antibiotic- resistant strains place Y. pestis at the top of the U.S. select agent list as a potential bioterrorism agent. Our long-term goal is to elucidate molecular mechanisms underlying the acute bacterial infectious process of Y. pestis . The more immediate objective is to identify and evaluate new and existing antigens of Y. pestis to develop a new generation plague vaccine. The complete genome sequence of Y. pestis is now known. Four aims are proposed. In Aim 1 we will prepare a Braun/murein lipoprotein (lpp)-minus mutant of Y. pestis , based on our recent data that the patented Ipp isogenic mutants of Salmonella Typhimurium and of Y pseudotuberculosis are avirulent in mice and provide protection against challenge with the wild-type bacterium. We will examine these mutants for immunological responses in mice to develop a live attenuated Y. pestis vaccine or use Y. pseudotuberculosis as a carrier for Y. pestis antigens. Aim 2 will identify differentially or exclusively expressed genes (potentially virulence-associated) of Y. pestis by genomics and proteomics to evaluate new antigens for use in a recombinant subunit plague vaccine. Aim 3 will examine the virulence potential of selected in vivo-expressed genes of Y. pestis by developing isogenic mutants and evaluating them for lethality in a mouse model and assess selected antigens' ability to provide immunity against Y. pestis challenge. Aim 4 will examine potential use of the Ipp-minus mutants of Y. pseudotuberculosis and S. Typhimurium as carriers to deliver Y. pestis antigens by expressing selected genes either from a plasmid under an inducible promoter and/or chromosome of attenuated Y. pseudotuberculosis/S. Typhimurium or by DMA vaccination. Alternatively, a vaccine strain of S. Typhi (Ty21a) could also be used. We believe these multiple approaches will identify candidate antigens for a use in a new, efficacious plague vaccine.