Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display

使用噬菌体 T4 展示多价鼠疫、炭疽疫苗

• 批准号: 7783779
• 负责人: Venigalla B. Rao
• 金额: $ 25.2万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-11 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7783779
• 关键词: Adjuvant; Aerosols; Anthrax Vaccines; Anthrax disease; Antigens; Bacillus anthracis; Bacteriophage T4; Binding; Biochemical; Breathing; Calcium; Capsid; Capsid Proteins; Categories; Complex; Drug Formulations; Escherichia coli; Goals; Immune response; In Vitro; Length; Lethal Dose 50; Macaca mulatta; Modeling; Molecular Genetics; Needles; Oryctolagus cuniculus; Plague; Plague Vaccine; Pneumonic Plague; Rattus norvegicus; Reproduction spores; Route; System; Testing; Type III Secretion System Pathway; Vaccines; Virulent; Yersinia; Yersinia pestis; anthrax lethal factor; anthrax toxin; biodefense; density; design; immunogenic; immunogenicity; nanoparticle; neutralizing antibody; next generation; nonhuman primate; novel; particle; pathogen; preclinical study; response; skin patch; vaccine efficacy

DESCRIPTION (provided by applicant): We hypothesize that in vitro assembly of antigens (Ags) on bacteriophage T4 can be developed as a common platform for construction of next generation multivalent biodefense vaccines. The major goal of this proposal is to test this hypothesis and develop trivalent plague and multivalent plague-anthrax vaccines that can confer protection against Bacillus anthracis and Yersinia pestis, two Category A biodefense pathogens. We have developed a defined in vitro system to array large foreign Ags at high density on phage T4 nanoparticle (120 x 86 nm). Full-length anthrax toxins, as large as 90kDa, are fused to the two non-essential phage T4 outer capsid proteins, Soc (small outer capsid protein; 870 copies) and Hoc (highly antigenic outer capsid protein; 155 copies), over-expressed in E. coli, purified to homogeneity, and displayed on hoc_soc_ capsid through in vitro binding. Multiple Ags and large hetero-oligomeric complexes can be displayed to saturation and the copy number can be controlled. The spatially exposed and symmetrically arrayed T4-rPA particles are highly immunogenic without any adjuvant, eliciting strong PA-specific and lethal toxin neutralizing Antibody (Ab) titers, conferring complete protection to rabbits against 100 LD50 B. anthracis Ames spore challenge. The best vaccine formulations are being tested in a preclinical trial using rhesus macaques. Combining our biochemical, molecular genetic, structural, and aerosol challenge expertise, strategies are designed to develop a novel trivalent T4-plague vaccine comprised of three Yersinia Ags, the capsular Ag, Caf1, the low calcium response V Ag, LcrV, and the Yersinia secretory complex factor, YscF. Inclusion of YscF, a highly conserved needle-forming subunit of type III secretion system that is essential to all virulent Yersinia species, is expected to generate a superior and broadly effective trivalent plague vaccine. The structural disposition and copy number of the capsid-bound plague Ags will be optimized. Combinations of T4-plague Ags, adjuvants, route of delivery (including skin patch), will be tested for Ab and cellular immune responses and protection against challenge with the capsular strain Y. pestis CO92, one of the most lethal plague strains. Efficacy of protection will be assessed in a new Brown Norway rat aerosol challenge model. The best T4-anthrax and T4-plague vaccines will be combined to create a novel multivalent anthrax-plague vaccine. The efficacy of this vaccine for protection against both inhalation anthrax and pneumonic plague will be tested in rhesus macaque aerosol challenge in model.

描述（由申请人提供）：我们假设噬菌体T4上的抗原（AGS）体外组装可以作为建造下一代多价生物化疫苗的通用平台。该提案的主要目的是检验这一假设，并发展三价鼠疫和多价疫苗疫苗，这些疫苗可以赋予炭疽芽孢杆菌和耶尔森氏菌的保护，这是两类A类生物固定病原体。我们已经开发了一个定义的体外系统，以在噬菌体T4纳米粒子（120 x 86 nm）上以高密度的大型外国AG阵列。 Full-length anthrax toxins, as large as 90kDa, are fused to the two non-essential phage T4 outer capsid proteins, Soc (small outer capsid protein; 870 copies) and Hoc (highly antigenic outer capsid protein; 155 copies), over-expressed in E. coli, purified to homogeneity, and displayed on hoc_soc_ capsid through in vitro binding.多个AGS和大型异寡聚体配合物可以显示为饱和，并且可以控制拷贝数。空间暴露和对称阵列的T4-RPA颗粒具有高度免疫原性，而没有任何辅助物，引起了强烈的PA特异性和致命的毒素中和抗体（AB）滴度（AB）滴度，从而为兔子提供了针对100 LD50的B. Anthracis Ames Spore挑战的兔子的完全保护。最好的疫苗制剂正在使用恒河猕猴的临床前试验中测试。结合我们的生化，分子遗传，结构和气雾剂挑战专业知识，旨在开发一种新型的三价T4斑块疫苗，由三个Yersinia ags组成，Capsular Ag，Caf1，Caf1，低钙反应V AG，LCRV，LCRV和Yersinia sermisinia sermisinia sermitolory Complextory复数因子，YSCF，YSCF，YSCF。包括YSCF是III型分泌系统的高度保守的针形成亚基，对所有有毒的Yersinia物种必不可少，预计将产生一种优越且广泛的三价瘟疫疫苗。将优化带束鼠疫AG的结构处置和拷贝数。 T4斑块AG，佐剂，递送途径（包括皮肤贴剂）的组合将测试AB和细胞免疫反应，并通过囊肿Y。PestisCO92（最致命的瘟疫菌株之一）进行挑战。保护功效将在新的棕色挪威大鼠气溶胶挑战模型中评估。最好将最佳的T4-Anthrax和T4-束疫苗组合起来，以创建一种新型的多价炭疽束疫苗。该疫苗可防止吸入炭疽和肺鼠疫的功效，将在模型中的恒河猴气溶胶挑战中进行测试。