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.

项目摘要 结核病（TB）是世界上最重要的疾病之一，并且是针对结核病的安全有效的疫苗。 病原体结核分枝杆菌（Mtb），其比目前仅部分可用的更有效 有效M.迫切需要牛菌株卡介苗（BCG）疫苗。一般认为 需要一种改进的BCG替代疫苗和一种有效的异源加强疫苗， 与结核病的斗争。该项目的目的是优化和进行先进的概念验证研究 在小动物和非人灵长类动物（NHP）中， 重组减毒单核细胞增生李斯特菌载体疫苗抗结核病。 活减毒重组单核细胞增生李斯特菌（rLm）疫苗提供了优于其他疫苗的主要优点。 加强疫苗的方法，包括佐剂中的蛋白质和病毒载体疫苗，就成本而言， 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 多缺失李斯特菌载体（Lm ΔactA ΔinlB prfA*），并证明表达四个关键基因的rLm疫苗 免疫保护性Mtb蛋白（rLmMtb 4Ag）当用作免疫保护剂时， 在小鼠中针对Mtb气溶胶攻击的初免-加强免疫接种策略中的异源加强疫苗， 豚鼠此外，通过第一代rLm载体递送免疫保护性Mtb蛋白更容易。 比通过重组病毒载体递送或与有效佐剂一起施用更有效。 本申请的目的是优化表达4 Mtb的Lm-载体疫苗的表达 抗原;将其抗原库扩展到六种抗原以提高其效力;并评估优化的 作为一种独立疫苗和作为一种疫苗的安全性、免疫原性和有效性的最终先导rLm疫苗候选物 在小鼠、豚鼠和非人灵长类动物（NHP）中给予BCG致敏动物异源加强疫苗 肺结核模型。我们将通过以下方式实现这一目标：a）优化以下的蛋白表达盒： rLmMtb 4Ag疫苗;系统评价其他新型Mtb抗原的免疫原性和在 小鼠，选择前两种抗原，随后构建rLmMtb 6 Ag先导疫苗候选物; B） 对优化的rLmMtb 6Ag先导候选疫苗进行全面的概念验证研究， 作为单独和异源加强疫苗在小鼠模型中的安全性、免疫原性和有效性 c）对先导rLmMtb 6Ag疫苗作为标准进行选定的概念验证研究， 单独和异源加强疫苗在豚鼠模型中的安全性、免疫原性和疗效 肺结核;以及d）由于Aeras需要NHP中的概念验证以使疫苗进入临床前开发- 评价了主要的rLmMtb 6Ag候选疫苗作为独立疫苗的安全性、免疫原性和 与Aeras、Bioqual和Tulane National Primate合作在NHP肺结核模型中的疗效 研究中心。