Immunotherapeutics and vaccines against anthrax, plague and tularemia

生物防御研究的重点应包括开发免疫治疗剂和对抗生物战剂的有效疫苗。细胞免疫和体液免疫应答在炭疽、鼠疫和兔热病的免疫中均起作用。Vy 2 V82 T细胞仅存在于灵长类动物中，占血液中总人γ 5 T细胞群的60-95%。我们和其他人已经证明，磷酸化抗原特异性Vy 2 V82 T细胞可以作为先天性和适应性免疫细胞发挥作用，并有助于对急性致命形式的结核病的适应性免疫。我们的新研究表明，B。炭疽杆菌确实可以产生磷酸化抗原刺激V γ 2 V 52 T细胞，并在随后感染产生磷酸化抗原的分枝杆菌后引发这些细胞的交叉反应性记忆型应答。重要的是，猴的磷酸化抗原HMBPP处理可以从总循环T细胞中的70%扩增V γ 2 V δ 2 T细胞<1% to >。HMBPP活化的V γ 2 V82 T细胞容易迁移到肺，并在猕猴中提供针对肺分枝杆菌感染的保护。我们假设磷酸化抗原HMBPP介导的V γ 2 V 52 T细胞的活化可以极大地增强对B的先天性和适应性免疫应答。anthracis，Y. pestis和F.土拉热，并赋予保护免受致命的吸入性炭疽，鼠疫和土拉菌病。我们进一步假设，由磷抗原和蛋白抗原组成的联合疫苗可以靶向4种免疫组分：Vy 2 V52 T细胞、CD 4 T辅助细胞、CD 8 T杀伤细胞和抗体，因此可以更有效地用于免疫和疫苗诱导的针对炭疽、鼠疫和兔热病的保护。为了便于测试这些假设，我们已经制定了HMBPP治疗方案的临床和免疫学方面，并开发了用于构建重组BCG和李斯特菌疫苗载体的疫苗平台技术。为了验证我们的假设，并最终开发出针对炭疽、鼠疫和兔热病的重组疫苗，我们将：评估HMBPP方案对非人灵长类动物致命性吸入性炭疽、鼠疫和兔热病的免疫效果。二.构建并鉴定表达B的重组卡介苗和重组减毒李斯特菌疫苗载体。炭疽病宾夕法尼亚州，Y.鼠疫F_1或F.土拉菌属三.比较口服和皮内免疫途径在接受表达B的重组BCG和李斯特菌疫苗载体的异源初免-加强免疫的猴子中疫苗引起的免疫应答。anthracis PA，Y. pestis F1或F.土拉菌属四.确定口服异源性初免-加强免疫接种重组卡介苗和李斯特菌疫苗载体表达PA，F1，或兔热病抗原是否可以赋予非人灵长类动物对致命的吸入性炭疽，鼠疫和兔热病的保护性免疫。

DESCRIPTION (provided by applicant): Biodefense research priorities should include development of immunotherapeutics and effective vaccines against biological warfare agents. Both cellular and humoral immune responses may play a role in immunity to anthrax, plague and tularemia. Vy2V82 T cells exist only in primates and constitute 60-95% of total human y5 T cell population in the blood. We and others have demonstrated that phosphoantigen-specific Vy2V82 T cells can function as both innate and adaptive immune cells, and contribute to adaptive immunity to acutely fatal form of tuberculosis. Our new studies indicate that B. anthracis can indeed produce phosphoantigen stimulating Vy2V52 T cells and prime these cells for cross-reactive memory-type response after subsequent infection with phosphoantigen-producing mycobacteria. Importantly, phosphoantigen HMBPP treatment of monkeys can expand Vy2V52 T cells from <1% to > 70% in total circulating T cells. HMBPP-activated Vy2V82 T cells readily migrate to the lung and confer protection against pulmonary mycobacterial infection in macaques. We hypothesize that phosphoantigen HMBPP-mediated activation of Vy2V52 T cells can greatly boost innate and adaptive immune responses to B. anthracis, Y. pestis and F. tularensis, and confer protection against fatal inhalational anthrax, plague and tularemia. We further hypothesize that a combined vaccine comprised of both phosphoantigen and protein antigen can target 4 immune components: Vy2V52 T cells, CD4 T helpers, CDS T killers, and antibodies, and therefore can be more efficient for immunization and vaccine-induced protection against anthrax, plague and tularemia. To facilitate testing these hypotheses, we have worked out clinical and immunological aspects of HMBPP treatment regimens, and developed vaccine platform technology for constructing recombinant BCG and Listeria vaccine vectors. To test our hypothesis and ultimately develop recombinant vaccines against anthrax, plague and tularemia, we will: I. Assess HMBPP regimens for immunotherapeutic effects on fatal inhalational anthrax, plague, and tularemia in nonhuman primates. II. Construct and characterize recombinant BCG and recombinant attenuated Listeria vaccine vectors expressing B. anthrax PA, Y. pestis F1 or F. tularensis Ag. III. Compare oral and intradermal immunization routes for vaccine-elicited immune responses in monkeys that receive heterologous prime-boost vaccination with recombinant BCG and Listeria vaccine vectors expressing B. anthracis PA, Y. pestis F1, or F. tularensis Ag. IV. Determine whether oral heterologous prime-boost vaccination with recombinant BCG and Listeria vaccine vectors expressing PA, F1, or tularemia Ag can confer protective immunity against fatal inhalational anthrax, plague and tularemia in nonhuman primates.