Identification of New Antigens for a Plague Vaccine

鼠疫疫苗新抗原的鉴定

• 批准号: 6905101
• 负责人: ASHOK K CHOPRA
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6905101
• 关键词: HeLa cells; Yersinia; Yersinia pestis; Yersinia pestis disease; attenuated microorganism; bacterial antigens; bacterial vaccines; biotechnology; bioterrorism /chemical warfare; cytokine; emerging infectious disease; enzyme linked immunosorbent assay; gene expression; immune response; laboratory mouse; live vaccine; microarray technology; molecular cloning; northern blottings; polymerase chain reaction; protein biosynthesis; proteomics; vaccine development; vector vaccine; virulence

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.

描述（由申请人提供）：鼠疫耶尔森氏菌是急性鼠疫和肺鼠疫的病原体，也是人类经历过的最具破坏性的流行病细菌之一，现已被世界卫生组织列为重新出现的人类病原体。传染性的潜力、有效疫苗的缺乏以及多种抗生素耐药菌株的出现，使得鼠疫耶尔森菌成为美国潜在生物恐怖主义病原体的首选病原体。我们的长期目标是阐明鼠疫耶尔森菌急性细菌感染过程的分子机制。更直接的目标是识别和评估新的和现有的鼠疫耶尔森抗原，以开发新一代鼠疫疫苗。鼠疫耶尔森氏菌的完整基因组序列现已已知。提出了四个目标。在目标 1 中，我们将制备鼠疫耶尔森氏菌的 Braun/murein 脂蛋白 (lpp)-minus 突变体，根据我们最近的数据，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是无毒的，并提供针对鼠疫杆菌攻击的保护。野生型细菌。我们将检查这些突变体在小鼠中的免疫反应，以开发鼠疫耶尔森氏菌减毒活疫苗或使用假结核耶尔森氏菌作为鼠疫耶尔森氏菌抗原的载体。目标 2 将通过基因组学和蛋白质组学鉴定鼠疫耶尔森氏菌的差异或排他性表达基因（可能与毒力相关），以评估用于重组亚单位鼠疫疫苗的新抗原。目标 3 将通过开发同基因突变体并评估它们在小鼠模型中的致死率，来检查选定的鼠疫耶尔森氏菌体内表达基因的毒力潜力，并评估选定的抗原提供针对鼠疫耶尔森氏菌攻击的免疫力的能力。目标 4 将检查假结核耶尔森氏菌和鼠伤寒沙门氏菌的 Ipp-minus 突变体作为载体传递鼠疫耶尔森氏菌抗原的潜在用途，方法是表达来自诱导型启动子和/或减毒假结核耶尔森氏菌/染色体下的质粒的选定基因。 S。鼠伤寒或通过 DMA 疫苗接种。或者，也可以使用伤寒沙门氏菌 (Ty21a) 疫苗株。我们相信，这些多种方法将鉴定出用于新型有效鼠疫疫苗的候选抗原。