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）：三种大流行病的耶尔森尼亚鼠尾草导致了大约2亿张鼠疫死亡，并且在世界范围内仍然流行，导致零星的感染。由于无法过腐生生命及其在许多啮齿动物种群中的住所，因此瘟疫是最担心的对人的人畜共患病疾病的恐惧之一。至少在800年前，瘟疫开始被用作生物武器，如今已成为最可能的生物威胁之一。由于这些考虑因素，我们建议：（i）构建和评估重组的减毒伤寒沙门氏菌疫苗（RASV）合成Y. pestis k1m K1M抗原在体内口服后体内的抗原，以鉴定抗原，以鉴定抗原刺激性免疫性，以挑战毒性的pestis Y. Pestis k1m。 （ii）用RPOS*表型构建和评估一种重组型Typhi ISP1820，以传递一种或多种保护性鼠疫杆菌Kim抗原。如果发现需要不同的抗原递送模式以诱导Y.柴油挑战的保护性免疫，我们将建造所有被认为是必要的重组疫苗，以作为鸡尾酒给予。 （iii）构建和评估一种活死的鼠疫疫苗疫苗，该疫苗将引起对大多数（如果不是全部）鼠疫菌所用的蛋白质的免疫反应增强，以获取铁，重要的毒力属性，并且在由于阿拉伯蛋白被调制的延迟延迟的十个延迟分裂系统后，没有生存的生长。我们将使用缺乏70 kb lcr质粒的Y. pestis kim6+开始构造努力，并且在引入衰减突变后添加了LCR质粒。 我们还将开发我们的主文件，准备并充分表征候选疫苗主种子以稳定和安全，准备IRB批准的协议，提交必要的信息以获取IND，并执行任何其他工作，以安排在人类志愿者中对临床评估的最佳候选疫苗进行临床评估。