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2亿人死亡，目前仍在世界各地流行，导致零星感染。由于鼠疫不能过上嗜血的生活，并且在许多啮齿动物种群中存在，因此鼠疫是由专性动物-人类病原体引起的最可怕的人畜共患疾病之一。鼠疫杆菌至少在800年前开始被用作生物武器，今天是最有可能的生物威胁之一。基于这些考虑，我们提出：（i）构建和评价合成Y.鼠疫K1 M抗原，以鉴定刺激保护性免疫的抗原，以攻击强毒Y。鼠疫K1 M. (ii)重组减毒沙门氏菌的构建及鉴定。具有RpoS* 表型的伤寒ISP 1820以递送一种或多种保护性Y。鼠疫KIM抗原。如果发现诱导对Y的保护性免疫需要不同的抗原递送模式。鼠疫的挑战，我们将构建所有被认为必要的重组疫苗作为鸡尾酒施用。(iii)构建并评价活的减毒Y.鼠疫疫苗，将诱导增强免疫反应的大多数，如果不是全部，蛋白质所使用的耶尔森氏菌。鼠疫杆菌获得铁，一个重要的毒力属性，并表现出完全的生物遏制，由于阿拉伯糖调节的延迟细胞裂解系统，在十代生长后没有存活的细菌细胞。我们将与Y.鼠疫KIM 6+的Lcr质粒，并在引入减毒突变后加回Lcr质粒。 我们还将开发我们的主文件，准备和充分表征候选疫苗主种子的稳定性和安全性，准备和提交IRB批准的方案，提交获得IND所需的信息，并执行安排最佳候选疫苗在人类志愿者中进行临床评价所需的任何其他工作。