Advanced anthrax vaccine made with ISS DNA formulations

描述(申请人提供)：我们建议使用ISS开发一种改进的炭疽疫苗，该疫苗将是安全的，并将在一到两次免疫后提供免疫力。这种疫苗将由重组炭疽保护性抗原(RPA)与免疫刺激(ISS)寡核苷酸佐剂一起交付，并由一种新的凝集制剂增强。我们将比较目前处于临床试验中的ISS序列1018与我们更活跃的ISS序列C274，以确定最有效的疫苗配方。在新型凝聚载体中使用ISS，与目前使用吸附在明矾上的RPA的第二代方法或与目前获得许可的疫苗AVA相比，应能显著提高针对RPA产生的抗体反应的速度和幅度。该项目的具体目标是： 1.检测重组炭疽保护性抗原(RPA)在小鼠、兔和恒河猴体内与ISS凝集制剂联合使用前后的免疫原性；用PA特异性的ELISA和体外细胞培养试验评估血清中和炭疽毒素的能力。 2.用炭疽芽胞杆菌孢子攻击实验动物，观察1~2剂ISS凝集剂/RPA疫苗在体内对炭疽病的保护作用。 A.通过鼻腔给药保护小鼠免受炭疽芽胞的攻击。 B.用炭疽杆菌孢子气雾剂攻击后保护兔子免受吸入性炭疽病的影响。 C.用炭疽芽胞攻击后，恒河猴对吸入性炭疽的保护。 3.进行毒理学评估，并启动ISS凝聚/RPA疫苗的GMP生产，假定特定目标1-3取得积极结果。 由于后几个具体目标(目标3C和4)的成本较高，每个具体目标的活动将按顺序进行，并将在项目进入下一个目标之前确定可行性。假设个人特定目标取得成功，这项提案旨在生产一种可迅速进入临床测试的IND准备就绪的新炭疽疫苗。

DESCRIPTION (provided by applicant): We propose to use ISS to develop an improved anthrax vaccine that will be safe and will provide immunity after one or two immunizations. This vaccine will be compromised of recombinant anthrax protective antigen (rPA) delivered with an immunostimulatory (ISS) oligonucleotide adjuvant enhanced by a novel coacervate formulation. We will compare the ISS sequence currently in clinical trials, 1018, with our more active ISS sequence, C274 to identify the most potent vaccine formulation. The use of ISS in the novel coacervate carrier should provide dramatic enhancement of both the speed and magnitude of the antibody response developed against rPA compared to current second generation approaches using rPA adsorbed to alum or compared to the currently licensed vaccine, AVA. The specific aims for this project are: 1. To test the immunogenicity of recombinant anthrax protective antigen (rPA) with and without combination with ISS coacervate formulations in mice, rabbits, and rhesus macaques; sera will be evaluated by PA-specific ELISAs and for the capacity to neutralize the anthrax toxins using in vitro cell culture assays. 2. To establish the ability of 1-2 doses of the optimized ISS coacervate/rPA vaccine to protect experimental animals against anthrax in vivo following challenge with Bacillus anthracis spores. A. Protection of mice against challenge with B. anthracis spores administered by the intranasal route. B. Protection of rabbits against inhalation anthrax following aerosol challenge with B. anthracis spores. C. Protection of rhesus macaques against inhalation anthrax following challenge with B. anthracis spores. 3. To perform toxicological evaluation and to initiate GMP production of the ISS coacervate/rPA vaccine, assuming positive results in Specific Aims 1-3. Because of the high cost of the later Specific Aims (Aims 3C and 4), activities for each Specific Aim will be performed sequentially, and feasibility will be established before the project moves onto the next Aim. Assuming success in the individual Specific Aims, this proposal is intended to produce an IND-ready new anthrax vaccine that could rapidly move into clinical testing.