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: 8819513
• 负责人: Venigalla B. Rao
• 金额: $ 59.35万
• 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/8819513
• 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; Drug Formulations; Engineering; Future Generations; Genes; Goals; Grant; Head; Health; Immune response; Immunity; Lead; Legal patent; Lethal Dose 50; Macaca; Macaca mulatta; Modeling; Modification; Motor; Mucosal Immune Responses; Mucosal Immunity; Mus; Mutate; New Zealand; Oral; Oryctolagus cuniculus; Outcome; Plague; Plague Vaccine; Play; Pneumonic Plague; Population; Process; Production; Proteins; Publications; Published Comment; 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）：我们提出了一种使用噬菌体 T4 纳米颗粒递送平台来制造单剂量、多价生物防御疫苗的新方法。在两项非常成功的 U01 生物防御资助（17 篇出版物和 5 项专利）中，我们开发了一种 T4 保护性抗原 (PA) 炭疽疫苗，为兔子和恒河猴提供全面的保护，抵御炭疽杆菌孢子的 100 LD50 气溶胶攻击，以及 T4-F1mut-V（突变荚膜抗原 F1 和低钙反应 V 抗原）鼠疫疫苗，为小鼠和棕色挪威大鼠针对 5,000 LD50 鼠疫耶尔森氏菌 CO92 的鼻内或气溶胶攻击。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-鼠疫双疫苗，可以预防吸入性炭疽和肺鼠疫，ii）建立一个“即插即用”的 T4 疫苗输送平台，可以适应任何生物防御或新出现的病原体。 炭疽 PA 和鼠疫 F1mut-V 抗原将作为小外衣壳蛋白 Soc（870 个拷贝）的融合蛋白展示在 120 nm x 86 nm 噬菌体 T4 衣壳上。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 包装机包装在衣壳内。 DNA分子将诱导宿主产生抗原，持续数周至数月。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（目标 1）。将使用单剂量的这些制剂（不含佐剂）对小鼠进行免疫，并使用我们新开发的炭疽-鼠疫双重攻击模型筛选针对炭疽和鼠疫的保护作用。特别令人感兴趣的是外部含有抗原、内部含有 DNA 的“初免-加强”疫苗。通过不断表达抗原，蛋白质将充当“启动”，DNA充当“增强”，进而刺激有效的免疫反应。免疫学分析将量化结合抗体滴度、致命毒素中和滴度和细胞反应。将进一步评估最佳疫苗配方的反应持久性以及通过口服给药诱导粘膜免疫的能力（目标 2）。小鼠研究中的两种最佳配方将在第二种动物模型中进行测试，即用于治疗吸入性炭疽的新西兰白兔和用于治疗肺鼠疫的挪威棕色大鼠（目标 3）。最后，将通过用鼠疫耶尔森氏菌 CO92 和炭疽芽孢杆菌 Ames 孢子进行双重气溶胶攻击，在食蟹猴模型中评估它们的免疫原性和保护功效（目标 4）。 通过开发强大的疫苗输送平台、最先进的设施、与成熟的行业合作伙伴的合作伙伴关系以及一支实力互补的高水平研究人员团队，该提案将导致创建一种有效的炭疽-鼠疫双重疫苗这将对生物防御疫苗计划产生巨大影响。