An Oral Vaccine Against Multiple Biowarfare Agents

描述(由申请人提供)：背景：美国公民，特别是 军事人员，容易受到暴露于生物的威胁 战地特工。两种这样的细菌制剂，炭疽芽孢杆菌和弗朗西塞氏菌 图拉尔病毒很容易通过气雾化传播，导致高水平的 死亡率高，因此被认为是候选战剂。新的 需要针对这些和其他潜在战剂的疫苗，这些疫苗可以 易于管理，并提供高水平的防护 雾化生物武器。我们已经培育出一株鼠伤寒沙门氏菌。 (Delta-glnA Delta-glnH)具有许多使其具有吸引力的属性 一种减毒活疫苗的候选者。我们的假设是 该减毒鼠伤寒沙门氏菌株可作为单一口服疫苗。 传递多价抗原，并提供粘膜和系统保护 对雾化生物战剂的免疫力，特别是B。 炭疽菌和图拉氏镰刀菌。我们将开发特定的鼠伤寒沙门氏菌启动子 (例如，pmrH)驱动炭疽杆菌和图拉氏杆菌的高水平表达 淋巴组织内的抗原，从而产生足够的免疫力 只需一剂即可起效。 本项目的具体目标包括：(1)建设三角洲-GlnA 含pmrH启动子的Delta-glnH减毒鼠伤寒沙门氏菌 炭疽杆菌的驱动力表达。保护性抗原(PA)和图拉氏丝虫 FOPA和TUL4蛋白；(2)疫苗株的效力评价 (特定目标1)在免疫组织内表达异源抗原，并 激发适当的免疫反应；及(3)挑战接种疫苗的动物 用炭疽杆菌和图拉氏杆菌雾化吸入测定其疗效 疫苗株。我们的研究设计纳入了协作疫苗 根据专业知识，在圣安东尼奥的三个不同地点进行开发 在每个地点都能找到。表达炭疽杆菌的鼠伤寒沙门氏菌疫苗株 和图拉氏杆菌抗原将被构建并接种到动物体内 两名鼠伤寒沙门氏菌研究人员卡尔·克洛泽博士和约翰博士的实验室 来自德克萨斯大学健康科学中心的Gunn博士说。评价表 免疫组织内的抗原表达水平将在 布鲁克斯空军基地，作者：肯顿·罗曼博士。气雾化炭疽杆菌和F. 图拉尔人对接种疫苗动物的挑战研究将在 西南基金会生物安全4级(BSL-4)实验室 Jean Patterson博士指导下的生物医学研究。我们会的 利用这个高水平的生物遏制实验室来执行 证明这种疫苗方法的有效性所必需的气雾剂挑战。 相关性：单一口服疫苗的开发可以同时 提供对多种生物战剂的保护将是巨大的 有益于军事人员和其他暴露于 这些特工。如果这种疫苗策略被证明是成功的，额外的抗原 可以从相同的疫苗株中表达，提供了一种适应性和 保护性健康工具。

DESCRIPTION (provided by applicant): Background: U.S. citizens, particularly military personnel, are vulnerable to the threat of exposure to biological warfare agents. Two such bacterial agents, Bacillus anthracis and Francisella tularensis, can be easily spread by aerosolization causing a high level of mortality, and are therefore considered to be candidate warfare agents. New vaccines against these and other potential warfare agents are needed which can be easily administered and provide high levels of protection against aerosolized bio-weapons. We have developed a Salmonella typhimurium strain (delta-glnA delta-glnH) with a number of attributes that make it an attractive candidate for a live attenuated multivalent vaccine. Our hypothesis is that this attenuated S. typhimurium strain can be used as a single oral vaccine to deliver multivalent antigens and provide both mucosal and systemic protective immunity against aerosolized biological warfare agents, specifically B. anthracis and F. tularensis. We will exploit specific S. typhimurium promoters (e.g., pmrH) to drive high-level expression of B. anthracis and F. tularensis antigens within the lymphoid tissue, and thus generate a sufficient immune response with a single dose. The Specific Aims of this project entail: (1) Construction of delta-glnA delta-glnH attenuated S. typhimurium vaccine strains with the pmrH promoter driving expression of B. anthracis. Protective Antigen (PA) and F. tularensis FopA and TUL4 proteins; (2) Evaluation of the efficacy of vaccine strains (Specific Aim 1) to express heterologous antigens within immune tissue and elicit an appropriate immune response; and (3) Challenge vaccinated animals with aerosolized B. anthracis and F. tularensis to determine efficacy of the vaccine strains. Our Study Design incorporates collaborative vaccine development at three different sites in San Antonio, based upon the expertise found at each site. The S. typhimurium vaccine strains expressing B. anthracis and F. tularensis antigens will be constructed and inoculated into animals in the laboratories of two S. typhimurium researchers, Drs. Karl Klose and John Gunn, at the University of Texas Health Science Center. The evaluation of levels of antigen expression within immune tissue will be carried out at the Brooks Air Force Base by Dr. Kenton Lohman. Aerosolized B. anthracis and F. tularensis challenge studies of vaccinated animals will take place in the Biosafety Level 4 (BSL-4) laboratory at the Southwest Foundation for Biomedical Research under the guidance of Dr. Jean Patterson. We will be taking advantage of this high-level biocontainment laboratory to perform the aerosol challenges necessary to prove the efficacy of this vaccine approach. Relevance: The development of a single oral vaccine that can simultaneously provide protection against multiple bio-warfare agents would be of tremendous benefit to the health of military personnel and other citizens exposed to these agents. If this vaccine strategy proves successful, additional antigens can be expressed from the same vaccine strain, offering an adaptive and protective health tool.