A Multicomponent Anthrax Vaccine using Phage T4 Display

描述（由申请人提供）：本申请的目的是开发一种多组分炭疽疫苗，该疫苗易于给药，诱导持久的高抗体滴度，并提供针对炭疽芽孢杆菌感染的保护。三个新的平台技术，无针皮肤贴片经皮免疫（TCI），噬菌体T4多组分展示，脂质体和乳剂佐剂配方，将承担开发一个有效的炭疽疫苗。这项研究将由两个互补的实验室进行，一个在T4噬菌体颗粒的遗传学和制造以及T4表面蛋白质的表达方面非常熟练，另一个在TCI，免疫原性脂质体，乳剂，疫苗制剂和疫苗临床试验方面具有广泛的经验，与第三个实验室密切合作，该实验室是炭疽毒素生物学和生产的领导者。所有三种炭疽毒素组分，保护性抗原（PA）、致死因子（LF）和水肿因子（EF）的突变形式将表达为单个蛋白质或表达为噬菌体T4的高抗原性外衣壳蛋白（Hoc）的N-末端融合物。将具有N-末端六组氨酸标签的毒素-Hoc融合蛋白大量纯化并使用体外组装系统加载到噬菌体T4二十面体表面上。将使用小鼠模型在平行追踪中评价可溶性蛋白或T4展示抗原与脂质体和乳剂佐剂制剂组合的TCI和肌内（IM）递送途径，以产生保护性抗体滴度。将用（i）炭疽毒素、（ii）Sterne菌株和（iii）艾姆斯菌株攻击免疫小鼠，以确定疫苗的效力。将在豚鼠模型中测试诱导小鼠对毒性炭疽芽孢杆菌攻击的保护作用的最佳组合。将对免疫应答进行表征，并进行炭疽毒力菌株的攻击实验，以选择诱导持久免疫的最佳疫苗产品。最有希望的炭疽疫苗候选者将在非人灵长类动物模型中进行测试，除了表征免疫应答外，免疫猕猴将用炭疽芽孢杆菌的雾化孢子进行攻击。将评估保护持续时间以及病理变化。

DESCRIPTION (provided by applicant): The goal of this application is to develop a multicomponent anthrax vaccine that can be easily administered, induces Iong-lasting high antibody titers, and provides protection against Bacillus anthracis infection. Three novel platform technologies, needle-free skin patch transcutaneous immunization (TCI), phage T4 multicomponent display, and liposome and emulsion adjuvant formulations, will be brought to bear on developing an efficacious anthrax vaccine. The research wilt be carried out by two complementary laboratories, one highly skilled in the genetics and manufacture of T4 bacteriophage particles and expression of proteins on the surface of T4, and the other with broad experience with TCI, immunogenic liposomes, emulsions, vaccine formulations, and vaccine clinical trials, in close collaboration with a third laboratory that is a leader in anthrax toxin biology and production. Mutant forms of all three anthrax toxin components, protective antigen (PA), lethal factor (LF) and edema factor (EF), will be expressed either as individual proteins or as N-terminal fusions of the highly antigenic outer capsid protein (Hoc) of bacteriophage T4. The toxin-Hoc fusion proteins with an N-terminal hexa-histidine tag will be purified in large quantities and loaded onto the phage T4 icosahedral surface using an in vitro assembly system. The TCI and intramuscular (IM) routes of delivery for combinations of soluble proteins or T4 displayed antigens with liposome and emulsion adjuvant formulations will be evaluated in parallel tracks, using the mouse model, for generation of protective antibody titers. The immunized mice will be challenged with (i) anthrax toxin, (ii) Sterne strain, and (iii) Ames strain, to determine the efficacy of the vaccines. The best combinations that induce protection in mice against challenge with virulent Bacillus anthracis will be tested in a guinea pig model. Immune responses will be characterized, and challenge experiments with the virulent anthrax strain will be performed to select the best vaccine product(s) that induce long-lasting immunity. The most promising anthrax vaccine candidates will be tested in a nonhuman primate model, in addition to characterization of immune responses, the immunized macaques will be challenged with the aerosolized spores of Bacillus anthracis. The duration of protection as well as pathological changes will be assessed.