Attenuated Live and Recombinant Yersinia pestis Vaccines

鼠疫耶尔森氏菌减毒活疫苗和重组疫苗

• 批准号: 7034308
• 负责人: ROY CURTISS III
• 金额: $ 54万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7034308
• 关键词: Salmonella typhimurium; Yersinia pestis; antigen antibody reaction; attenuated microorganism; bacterial toxins; bacterial vaccines; biotechnology; bioterrorism /chemical warfare; drug delivery systems; enzyme linked immunosorbent assay; flow cytometry; laboratory mouse; live vaccine; microorganism culture; molecular cloning; nucleic acid sequence; phase contrast microscopy; polymerase chain reaction; recombinant proteins; site directed mutagenesis; southern blotting; transmission electron microscopy; vaccine development; vaccine evaluation; vector vaccine; virulence

DESCRIPTION (provided by applicant): Yersinia pestis, in three pandemics, resulted in some 200 million plague deaths and is still endemic throughout the world resulting in sporadic infections. Due to its inability to lead a saprophytic life and its residence in many rodent populations, plague is one of the most feared of zoonotic diseases caused by an obligate animal-human pathogen. The plague bacillus began to be used as a biological weapon at least 800 years ago and is today one of the more likely biological threats. Because of these considerations, we propose to: (i) Construct and evaluate recombinant attenuated Salmonella typhimurium vaccines (RASV) synthesizing Y. pestis K1M antigens in vivo after oral immunization of mice to identify antigens that stimulate protective immunity to challenge with virulent Y. pestis K1M. (ii) Construct and evaluate a recombinant attenuated S. typhi ISP1820 with an RpoS* phenotype to deliver one or more protective Y. pestis KIM antigens. If it is found that different antigen delivery modes are required to induce protective immunity to Y. pestis challenge, we will construct all deemed necessary recombinant vaccines to be administered as a cocktail. (iii) Construct and evaluate a live attenuated Y. pestis vaccine that will induce enhanced immune responses to most, if not all, proteins used by Y. pestis to acquire iron, an important virulence attribute, and exhibit complete biological containment with no surviving bacterial cells after ten generations of growth due to an arabinose-regulated delayed cell lysis system. We will initiate our construction endeavors with Y. pestis KIM6+ that lacks the 70 kb Lcr plasmid and is totally avirulent and add back the Lcr plasmid after introducing attenuating mutations. We will also develop our Master File, prepare and fully characterize candidate vaccine Master Seeds for stability and safety, prepare and submit protocols for IRB approvals, submit information necessary to obtain INDs, and perform any other work needed to arrange that the best candidate vaccines be clinically evaluated in human volunteers.

描述（由申请人提供）：鼠疫耶尔森氏菌在三次大流行中导致约 2 亿人鼠疫死亡，并且仍然在世界范围内流行，导致零星感染。由于鼠疫无法过腐生生活，并且存在于许多啮齿类动物种群中，因此鼠疫是由动物-人类专性病原体引起的最令人恐惧的人畜共患疾病之一。鼠疫杆菌至少在 800 年前就开始被用作生物武器，如今已成为最有可能的生物威胁之一。出于这些考虑，我们建议：（i）构建和评估重组减毒鼠伤寒沙门氏菌疫苗（RASV），在小鼠口服免疫后体内合成鼠疫耶尔森氏菌 K1M 抗原，以鉴定刺激保护性免疫以抵抗强毒鼠疫耶尔森氏菌攻击的抗原。 K1M。 (ii)构建并评估具有RpoS*表型的重组减毒伤寒沙门氏菌ISP1820，以递送一种或多种保护性鼠疫耶尔森氏菌KIM抗原。如果发现需要不同的抗原递送模式来诱导针对鼠疫耶尔森氏菌攻击的保护性免疫，我们将构建所​​有认为必要的重组疫苗，以混合物的形式施用。 (iii) 构建并评估鼠疫耶尔森氏菌减毒活疫苗，该疫苗将诱导对鼠疫耶尔森氏菌用于获取铁（一种重要的毒力属性）的大多数（如果不是全部）蛋白质的增强免疫反应，并表现出完全的生物遏制，没有存活的细菌由于阿拉伯糖调节的延迟细胞裂解系统，细胞经过十代生长。我们将用缺乏 70 kb Lcr 质粒且完全无毒的鼠疫耶尔森氏菌 KIM6+ 开始我们的构建工作，并在引入减毒突变后添加回 Lcr 质粒。 我们还将开发我们的主文件，准备并充分描述候选疫苗主种子的稳定性和安全性，准备和提交 IRB 批准方案，提交获得 IND 所需的信息，并执行安排最佳候选疫苗上市所需的任何其他工作。在人类志愿者中进行临床评估。