Single Dose, Multivalent, Anthrax Plague Vaccines using Bacteriophage T4 Nanopart

使用噬菌体 T4 Nanopart 的单剂量、多价炭疽鼠疫疫苗

• 批准号: 9000614
• 负责人: Venigalla B. Rao
• 金额: $ 63.63万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-15 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9000614
• 关键词: Adjuvant; Aerosols; Animal Model; Anthrax Attack; Anthrax Vaccines; Anthrax disease; Antibodies; Antigens; Bacillus anthracis; Bacillus anthracis spore; Bacteriophage T4; Binding; Bioterrorism; Breathing; CMV promoter; Calcium; Capsid; Capsid Proteins; Chimeric Proteins; Collaborations; DNA; DNA Packaging; DNA delivery; Development; Dose; Engineering; Formulation; Future Generations; Genes; Goals; Grant; Head; Health; Immune response; Immunity; Lead; Legal patent; Lethal Dose 50; Macaca; Macaca mulatta; Modeling; Motor; Mucosal Immune Responses; Mucosal Immunity; Mus; Mutate; New Zealand; Oral; Oryctolagus cuniculus; Plague; Plague Vaccine; Play; Pneumonic Plague; Population; Process; Production; Proteins; Publications; Rattus; Rattus norvegicus; Reproduction spores; Research Personnel; Resources; Series; Surface; System; Technology; Testing; Translating; Vaccine Antigen; Vaccine Design; Vaccines; Yersinia pestis; anthrax lethal factor; anthrax protective factor; base; biodefense; biothreat; cell mediated immune response; density; design; immunogenicity; industry partner; interest; manufacturing process; multidisciplinary; nanoparticle; nonhuman primate; novel; novel strategies; particle; pathogen; programs; protective efficacy; response; success; vaccine delivery; vaccine development; vaccine efficacy; Adjuvant; Aerosols; Animal Model; Anthrax Attack; Anthrax Vaccines; Anthrax disease; Antibodies; Antigens; Bacillus anthracis; Bacillus anthracis spore; Bacteriophage T4; Binding; Bioterrorism; Breathing; CMV promoter; Calcium; Capsid; Capsid Proteins; Chimeric Proteins; Collaborations; DNA; DNA Packaging; DNA delivery; Development; Dose; Engineering; Formulation; Future Generations; Genes; Goals; Grant; Head; Health; Immune response; Immunity; Lead; Legal patent; Lethal Dose 50; Macaca; Macaca mulatta; Modeling; Motor; Mucosal Immune Responses; Mucosal Immunity; Mus; Mutate; New Zealand; Oral; Oryctolagus cuniculus; Plague; Plague Vaccine; Play; Pneumonic Plague; Population; Process; Production; Proteins; Publications; Rattus; Rattus norvegicus; Reproduction spores; Research Personnel; Resources; Series; Surface; System; Technology; Testing; Translating; Vaccine Antigen; Vaccine Design; Vaccines; Yersinia pestis; anthrax lethal factor; anthrax protective factor; base; biodefense; biothreat; cell mediated immune response; density; design; immunogenicity; industry partner; interest; manufacturing process; multidisciplinary; nanoparticle; nonhuman primate; novel; novel strategies; particle; pathogen; programs; protective efficacy; response; success; vaccine delivery; vaccine development; vaccine efficacy

DESCRIPTION (provided by applicant): We propose a novel approach to create single dose, multivalent, biodefense vaccines, using bacteriophage T4 nanoparticle delivery platform. In the two highly successful U01 biodefense grants (17 publications and 5 patents), we have developed a T4-protective antigen (PA) anthrax vaccine that provided complete protection to rabbits and rhesus macaques against 100 LD50 aerosol challenge of B. anthracis Ames spores, and a T4-F1mut-V (mutated capsular antigen F1 and low calcium response V antigen) plague vaccine that provided complete protection to mice and Brown Norway rats against intranasal or aerosol challenge with 5,000 LD50 of Yersinia pestis CO92. Building on these successes, this proposal will achieve two major goals: i) develop a single dose anthrax-plague dual vaccine that can protect against both inhalation anthrax and pneumonic plague, and ii) establish a "plug and play" T4 vaccine delivery platform that can be adapted to any biodefense or emerging pathogen. The anthrax PA and plague F1mut-V antigens will be displayed on the 120nm x 86nm phage T4 capsid shell, as fusion proteins of the small outer capsid protein, Soc (870 copies). The genes corresponding to these antigens will be cloned under the control of a strong CMV promoter and packaged inside the capsid, using the phage T4 DNA packaging machine. The DNA molecules will induce production of antigens in the host for weeks to months. A series of nanoparticle formulations will be prepared containing PA and F1mut-V antigens, genes, or both (Aim 1). Mice will be immunized with a single dose of these formulations, without an adjuvant, and screened for protection against both anthrax and plague, using our newly developed anthrax- plague double challenged model. Of particular interest are the "prime-boost" vaccines containing antigen outside and DNA inside. The protein would act as "prime" and the DNA as "boost," by continually expressing the antigens, and, in turn, stimulating potent immune responses. Immunological analyses will quantify binding antibody titers, lethal toxin neutralization titers, and cellular responses. The best vaccine formulations will be further evaluated for durability of the responses as well as the ability to induce mucosal immunity by oral delivery (Aim 2). The two best formulations from the mouse studies will be tested in a second animal model, New Zealand white rabbit for inhalation anthrax and Brown Norway rat for pneumonic plague (Aim 3). Finally, their immunogenicity and protective efficacy will be evaluated in the cynomolgus macaque model by double aerosol challenge with Y. pestis CO92 followed by Ames spores of B. anthracis (Aim 4). Synergized by the development of a robust vaccine delivery platform, state of the art facilities, partnership with an established industry partner, and a team of highly accomplished investigators with complementary strengths, this proposal would lead to the creation of an efficacious anthrax-plague dual vaccine that will have tremendous impact on the biodefense vaccine program.

描述（由申请人提供）：我们提出了一种新的方法，使用噬菌体T4纳米粒子递送平台创建单剂量，多价，生物纤维疫苗。 In the two highly successful U01 biodefense grants (17 publications and 5 patents), we have developed a T4-protective antigen (PA) anthrax vaccine that provided complete protection to rabbits and rhesus macaques against 100 LD50 aerosol challenge of B. anthracis Ames spores, and a T4-F1mut-V (mutated capsular antigen F1 and low calcium response V antigen) plague vaccine这为小鼠和棕色挪威大鼠提供了完全保护，以防止鼻内或气溶胶挑战，其中有5,000 ld50耶尔森氏菌CO92。在这些成功的基础上，该提案将实现两个主要目标：i）开发一种单剂量炭疽质量双疫苗，可以防止吸入炭疽和肺炎瘟疫，以及ii）建立一个“插头和游戏” T4疫苗交付平台，该平台可以适应任何生物粘液剂或任何生物疾病或出现的病原体。 炭疽PA和鼠疫F1MUT-V抗原将显示在120nm x 86nm Phage T4 Capsid壳上，作为小型外皮蛋白的融合蛋白，SOC（870份）。与这些抗原相对应的基因将在强CMV启动子的控制下克隆，并使用噬菌体T4 DNA包装机在衣壳内包装。 DNA分子将在宿主中诱导抗原的产生数周到数月。将制备一系列含有PA和F1MUT-V抗原，基因或两者的纳米颗粒制剂（AIM 1）。小鼠将使用我们新开发的炭疽病双重挑战的模型，将小鼠单一剂量的这些配方进行免疫，并在没有辅助的情况下进行筛选，以防止炭疽和瘟疫。特别令人感兴趣的是包含外部抗原和内部DNA的“原始促进剂”疫苗。该蛋白质将通过不断表达抗原，然后刺激有效的免疫反应来充当“ Prime”和DNA作为“增强”。免疫学分析将量化结合抗体滴度，致命的毒素中和滴度和细胞反应。最好的疫苗配方将进一步评估响应的耐用性以及通过口服递送诱导粘膜免疫的能力（AIM 2）。小鼠研究的两种最佳配方将在第二种动物模型，新西兰白兔子吸入炭疽和棕色挪威大鼠的肺炎鼠疫（AIM 3）中进行测试（AIM 3）。最后，将通过双气溶胶挑战在cynomolgus猕猴模型中评估它们的免疫原性和保护效率。 通过开发强大的疫苗输送平台，最先进的设施，与既定的行业合作伙伴的合作伙伴关系以及具有互补优势的高度成就的调查员团队的协同作用，该提案将导致有效的炭疽质质量双疫苗会对生物粘土式疫苗计划产生巨大影响。