Optimization and Advanced Proof-of-Concept Studies of a Listeria-vectored Multi-Antigenic Vaccine against Tuberculosis

李斯特菌载体多抗原结核疫苗的优化和高级概念验证研究

• 批准号: 10308053
• 负责人: MARCUS AARON HORWITZ
• 金额: $ 120.07万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10308053
• 关键词: Acquired Immunodeficiency Syndrome; Adjuvant; Aerosols; Affect; Amino Acid Substitution; Animals; Antigens; Attenuated; BCG Live; BCG Vaccine; Bioterrorism; Cavia; Clinical Research; Collaborations; DNA cassette; Development; Disease; Dose; ELF3 gene; Evaluation; Francisella tularensis; Generations; Glycoproteins; Goals; Human; Immune response; Immunity; Immunization; Immunize; Laboratories; Lead; Listeria; Listeria monocytogenes; Modeling; Mus; Mycobacterium bovis; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Patients; Persons; Planet Earth; Primates; Proteins; Public Health; Pulmonary Tuberculosis; Recombinants; Research; Risk; Route; Safety; System; Testing; Tuberculosis; Tuberculosis Vaccines; Tularemia; Vaccinated; Vaccines; Virulence; Virulent; Virus; booster vaccine; cancer cell; cost; early childhood; efficacy study; fight against; guinea pig model; immunogenicity; improved; lead candidate; macrophage; mouse model; nonhuman primate; novel; pathogen; pathogenic bacteria; preclinical development; protein expression; recombinant viral vector; secretory protein; vaccination strategy; vaccine candidate; vaccine immunogenicity; vaccine safety; vector; vector vaccine

PROJECT SUMMARY Tuberculosis (TB) is one of the world's most important diseases, and a safe and effective vaccine against the causative agent Mycobacterium tuberculosis (Mtb) that is more potent than the currently available only partially effective M. bovis strain Bacille Calmette-Guérin (BCG) vaccine is sorely needed. It is generally acknowledged that both an improved replacement vaccine for BCG and a potent heterologous booster vaccine are needed in the fight against TB. The purpose of this project is to optimize and conduct advanced proof-of-concept studies in small animals and non-human primates (NHP) of a second-generation heterologous multiantigenic recombinant attenuated Listeria monocytogenes-vectored vaccine against TB. Live attenuated recombinant Listeria monocytogenes (rLm) vaccines offer major advantages over other approaches to booster vaccines, including protein in adjuvant and virus-vectored vaccines, in terms of cost, ease of manufacture, immunogenicity and efficacy. In preliminary studies, we have identified an improved multi-deletional Listeria vector (Lm ΔactA ΔinlB prfA*) and demonstrated that rLm vaccines expressing four key immunoprotective Mtb proteins (rLmMtb4Ag) substantially augment protective immunity when used as a heterologous booster vaccine in a prime-boost vaccination strategy against Mtb aerosol challenge in mice and guinea pigs. Moreover, delivering the immunoprotective Mtb protein via a first generation rLm vector was more efficacious than delivering it via a recombinant viral vector or administering it with a potent adjuvant. The goal of this application is to optimize expression of an Lm-vectored vaccine expressing 4 Mtb antigens; expand its antigen repertoire to six antigens to increase its potency; and to evaluate the optimized final lead rLm vaccine candidate for safety, immunogenicity and efficacy as a standalone vaccine and as a heterologous booster vaccine to BCG-primed animals in mouse, guinea pig, and non-human primate (NHP) models of pulmonary TB. We shall accomplish this goal by: a) Optimizing the protein expression cassette of rLmMtb4Ag vaccine; systematically evaluating additional novel Mtb antigens for immunogenicity and efficacy in mice, selecting the top two antigens, and subsequently constructing a rLmMtb6Ag lead vaccine candidate; b) Conducting comprehensive proof-of-concept studies of the optimized rLmMtb6Ag lead vaccine candidate for safety, immunogenicity, and efficacy as standalone and heterologous booster vaccine in the mouse model of pulmonary TB; c) Conducting selected proof-of-concept studies of the lead rLmMtb6Ag vaccine as a standa- lone and heterologous booster vaccine for safety, immunogenicity and efficacy in a guinea pig model of pulmonary TB; and d) as Aeras requires proof-of-concept in NHP for a vaccine to enter preclinical develop- ment, evaluating the lead rLmMtb6Ag candidate as a standalone vaccine for safety, immunogenicity and efficacy in a NHP model of pulmonary TB in collaboration with Aeras, Bioqual, and Tulane National Primate Research Centre.

项目总结 结核病是世界上最重要的疾病之一，是预防结核病的一种安全有效的疫苗。 结核分枝杆菌(Mtb)，比目前仅有的部分有效 目前迫切需要有效的牛分枝杆菌卡介苗疫苗。这是公认的 需要一种改进的卡介苗替代疫苗和一种有效的异种加强疫苗 与结核病作斗争。该项目的目的是优化和进行高级概念验证研究 在小动物和非人灵长类动物(NHP)中的第二代异源多抗原 结核重组单核细胞增多性李斯特氏菌载体疫苗。 与其他疫苗相比，减毒重组单核细胞增生性李斯特菌(RLM)活疫苗具有重大优势 加强疫苗的方法，包括佐剂和病毒载体疫苗中的蛋白质，在成本方面， 易于制造、免疫原性和功效。在初步研究中，我们发现了一种改进的 多缺失李斯特氏菌载体(LMΔActaΔin1B PrFA*)，并证明了表达四个关键基因的RLm疫苗 免疫保护性Mtb蛋白(RLmMtb4Ag)作为一种 异种强化疫苗在小鼠和小鼠中对抗结核分枝杆菌气溶胶攻击的加强免疫策略 豚鼠。此外，通过第一代RLM载体传递免疫保护性Mtb蛋白 比通过重组病毒载体传递或使用强有力的佐剂更有效。 本应用的目的是优化表达4Mtb的LM载体疫苗的表达 抗原；将其抗原库扩大到六种抗原，以增加其效力；并评估优化的 作为一种独立疫苗和作为一种 卡介苗免疫小鼠、豚鼠和非人灵长类动物的异种强化疫苗(NHP) 肺结核病模型。我们将通过以下方式实现这一目标：a)优化蛋白质表达盒 RLmMtb4Ag疫苗；系统评价新的Mtb抗原的免疫原性和免疫效果 小鼠，选择前两个抗原，随后构建rLmMtb6Ag铅疫苗候选；b) 对优化的rLmMtb6Ag候选疫苗进行全面的概念验证研究 在小鼠模型中作为单独和异种加强疫苗的安全性、免疫原性和有效性 肺结核病；c)作为一个标准，对主要的rLmMtb6Ag疫苗进行选定的概念验证研究- 孤立型和异种佐剂疫苗在豚鼠肺炎模型中的安全性、免疫原性和有效性 肺结核病；以及d)由于AERS需要在NHP中进行概念验证，疫苗才能进入临床前开发- 评估领先的rLmMtb6Ag候选疫苗作为独立疫苗的安全性、免疫原性和 与AERAS、BIOQUAL和杜兰国家灵长类动物合作的NHP肺结核病模型的疗效 研究中心。