Immunological characterization of rationally-designed vaccines against plague in mice and non-human primate models

合理设计的鼠疫疫苗和非人灵长类动物模型的免疫学特征

• 批准号: 10335231
• 负责人: ASHOK K CHOPRA
• 金额: $ 57.49万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10335231
• 关键词: Acyltransferase; African Green Monkey; Animals; Antibiotic Resistance; Antibodies; Antibody Formation; Antibody Response; Antibody titer measurement; Antigens; Attenuated; Attenuated Vaccines; Bacteria; Bacterial Antigens; Binding; Bite; Bubonic Plague; Calcium; Case Fatality Rates; Categories; Cells; Cellular Immunity; Centers for Disease Control and Prevention (U.S.); Cessation of life; China; Clinical; Clinical Research; Clinical Trials; Congo; Coronavirus; Country; Data; Democratic Republic of the Congo; Disease; Disease Outbreaks; Dose; Engineering; Etiology; Exposure to; FDA approved; Fatality rate; Fleas; Fright; Future; Generations; Genes; Genetic Engineering; Goals; Human; Immune response; Immunity; Immunization; Immunize; Immunoglobulin A; Immunoglobulin G; Immunologics; Infection; Injections; Interferon Type II; Interleukin-17; Intramuscular; Iron; Laboratories; Lauric Acids; Left; Legal patent; Lesion; Link; Lipopolysaccharides; Lipoproteins; Macaca; Macaca fascicularis; Madagascar; Military Personnel; Modeling; Mucosal Immunity; Mus; Nature; Nepal; Neutrophil Infiltration; Patient-Focused Outcomes; Patients; Peptide Hydrolases; Plague; Plague Vaccine; Plasminogen Activator; Pneumonic Plague; Quarantine; Rattus; Reporting; Resistance; Rodent; Rodent Model; Role; Serum; Signal Transduction; Site; Subunit Vaccines; Symptoms; T cell response; TLR2 gene; TLR4 gene; Testing; Th1 Cells; Toxicology; Type III Secretion System Pathway; Vaccinated; Vaccine Design; Vaccines; Variant; Virulence; Yersinia pestis; Zoonoses; arm; base; bioweapon; climate change; co-infection; combat; cytokine; design; emerging human pathogen; immunogenicity; innovation; mouse model; mutant; nonhuman primate; novel; novel coronavirus; pandemic disease; pathogen; pre-clinical; resistant strain; respiratory; response; translational approach; vaccine access; vaccine candidate; vaccine efficacy

ABSTRACT The increasing number of bubonic/pneumonic plague cases globally (2010-2018), including the U.S., with a ~18% case fatality rate may reflect climate changes and a rodent carrier range shift. The 2017-18 plague outbreak in Madagascar with ~2400 cases (>75% pneumonic) and ~9% causalities has led WHO (April 2018) to intensify the need for developing new generation subunit and live-attenuated plague vaccines. This need is exemplified by deadly plague cases in China (2019) and Congo (2020 with a 35% fatality rate). Y. pestis’ (Yp) ability to persist in dead hosts to resurge after years of silence, existence of antibiotic-resistant strains that occur naturally or have been intentionally developed, and no FDA-approved plague vaccine, is fearsome. Two- component subunit vaccines composed of capsular antigen F1 and a T3SS component and effector LcrV (low calcium response V antigen), which only generate a humoral immune response, provide variable protection in African green monkeys (AGM) and generate poor T cell responses in humans. Such vaccines will not be effective against Yp strains lacking F1 or possessing LcrV variants. Since the cellular immunity is also critical for protection, we focused first on identifying new virulence genes of Yp and then to delete them in combination to develop novel live-attenuated vaccine (LAV) strains. Two such LAVs were 100% attenuated in inducing bubonic/pneumonic plague in mice/rats and generated long-term humoral and cellular immune responses to provide 100% protection to rodents against developing plague. No clinical symptoms of the disease or histopathological lesions were noted either during immunization or when the vaccinated animals were subsequently exposed to Yp CO92 in a more stringent pneumonic plague model. Therefore, further immunological characterization of these mutants and their testing in higher animals, such as cynomolgus macaques (CM) and AGM, will provide a rationale for future clinical studies. There is a precedent for using a LAV against plague (EV76 strain) in humans. However, this vaccine is reactogenic, represents a spontaneous mutant, and causes disease in patients with over iron load. In Aim 1, we will demonstrate efficacy and immune responses of two vaccine candidates generated from Yp CO92 (biovar Orientalis) against other Yp biovars (Antiqua and Medievalis), the F1-minus mutant of CO92, and Yp CO92 with LcrV variants, in bubonic and pneumonic mouse models. Our data with the mutants indicated a role of IL-17 (a Th17 cytokine), Th1-IFN-γ, and antibodies, in protection. In Aim 2, we will study the mechanistic basis of this protection (one chosen mutant) by using RORt-/- mice, which lack Th17 cells, as well as IFN-γ and IgA k/o mice, to discern their links to neutrophil recruitment and mucosal immunity, to combat Yp infection in bubonic/pneumonic plague models. In Aim 3, CM and AGM will be used with one mutant to demonstrate its short- and long-term efficacy in causing bubonic and/or pneumonic plague as well as reactogenicity. The correlates of protective immunity will then be established. These innovative mechanistic/translational approaches will result in effective new generation plague vaccines.

抽象的 全球的泡沫/肺炎鼠疫病例的数量增加（2010-2018），包括美国 〜18％的病例死亡率可能反映了气候变化和啮齿动物载体范围的变化。 2017-18瘟疫 马达加斯加的爆发约为2400例（> 75％的肺炎）和约9％的因果关系，导致Who（2018年4月） 增强开发新一代亚基和活瘟疫疫苗的需求。这是 中国（2019）和刚果（2020年，死亡率为35％）以致命的瘟疫案例为例。 Y. Pestis（YP） 多年的沉默，存在抗生素抗性菌株后，能够坚持死亡的宿主恢复活力的能力 自然或有意开发，并且没有FDA批准的瘟疫疫苗，这是可怕的。二- 组件亚基疫苗由帽抗原F1和T3SS组件和效应子LCRV组成（低 钙反应v抗原）仅产生体液免疫响应，可在 非洲绿色猴子（AGM）并在人类中产生不良的T细胞反应。这样的疫苗将无效 针对缺乏F1或具有LCRV变体的YP菌株。由于细胞免疫对于 保护，我们首先专注于识别YP的新病毒基因，然后将其删除 开发新型的活体疫苗（LAV）菌株。在诱导的 小鼠/大鼠的气泡/肺鼠疫，并产生长期的体液和细胞免疫调查 为啮齿动物提供100％的保护，以防止发育瘟疫。没有疾病的临床症状或 在免疫期间或接种动物是在接种疫苗时都会发现组织病理病变 随后在更严格的肺鼠瘟疫模型中暴露于YP CO92。因此，进一步 这些突变体的免疫学表征及其在较高动物中的测试，例如cynomolgus 猕猴（CM）和AGM将为将来的临床研究提供理由。有一个先例 对人类的鼠疫（EV76菌株）的lav。但是，该疫苗是反应源性的，代表了一个赞助 突变体，并导致铁负荷过度的患者疾病。在AIM 1中，我们将证明效率和免疫力 由YP CO92（Biovar Orientalis）对其他YP Biovars产生的两个疫苗候选者的反应 （Antiqua和Medievalis），CO92的F1-Minus突变体，以及具有LCRV变体的YP CO92，在气泡和 肺小鼠模型。我们使用突变体的数据表明IL-17（Th17细胞因子），Th1-IFN-γ和 抗体，保护。在AIM 2中，我们将研究此保护的机械基础（一个选择突变体） 使用缺乏Th17细胞以及IFN-γ和Iga K/O小鼠的Rort - / - 小鼠辨别它们与中性粒细胞的联系 招募和粘膜免疫，以打击气泡/肺鼠疫模型中的YP感染。在AIM 3，CM ANGM将与一个突变体一起使用，以证明其短期和长期效率在引起泡沫和/或 肺鼠疫以及反应生成。然后将建立保护性免疫的相关性。 这些创新的机械/翻译方法将导致有效的新一代平台疫苗。