Generation of a polycistronic universal influenza virus vaccine based on rare species adenoviral vectors

基于稀有物种腺病毒载体的多顺反子通用流感病毒疫苗的制备

• 批准号: 9806521
• 负责人: Lynda Coughlan
• 金额: $ 25.43万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-06 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9806521
• 关键词: Adenovirus Vector; Adjuvant; Adult; Animal Model; Antibody Formation; Antigen Targeting; Antigens; B-Lymphocytes; Biological Assay; Bronchoalveolar Lavage; CD8B1 gene; Cavia; Cells; Cessation of life; Cleaved cell; Clinical; Clinical Protocols; Clinical Trials; Cold Chains; Combined Vaccines; Communicable Diseases; DNA; DNA cassette; Data; Elderly; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitopes; Escherichia coli; Ferrets; Flow Cytometry; Future; Generations; Genome; Healthcare Systems; Hemagglutinin; Human; Immune; Immune response; Immunity; Immunoglobulin A; Immunoglobulin G; Immunologics; In Vitro; Individual; Infant; Infection; Influenza; Influenza A virus; Intramuscular; Lead; Measures; Mediating; Membrane Glycoproteins; Methods; Modeling; Molecular; Mucous Membrane; Mus; Mutation; Neuraminidase; Nucleoproteins; Peptides; Performance; Phase; Phenotype; Plaque Assay; Play; Population; Process; Production; Proteins; Protocols documentation; Public Health; Regimen; Research; Role; S100 Proteins; Safety; Seasons; Serotyping; Serum; Spleen; Stains; T cell response; T-Lymphocyte; Tertiary Protein Structure; Time; Transgenes; Tumor Antigens; Ultracentrifugation; Vaccination; Vaccine Antigen; Vaccine Production; Vaccines; Viral; Viral Antigens; Viral Vector; Virus Diseases; antigen-specific T cells; base; cesium chloride; cross reactivity; cytokine; design; economic impact; egg; exosome; experimental study; extracellular vesicles; homologous recombination; immunogenic; immunogenicity; immunoregulation; improved; in vivo; influenza virus vaccine; influenzavirus; innovation; interest; medical schools; mortality; nano-exosomes; nanosized; novel; pandemic disease; pandemic preparedness; particle; pathogen; promoter; respiratory; response; scale up; seasonal influenza; universal influenza vaccine; vaccine effectiveness; vector; vector vaccine

SUMMARY: Influenza A viruses (IAVs) cause serious respiratory illness in humans, with ~250,000-500,000 deaths per year globally. In addition to seasonal epidemics, the ongoing pandemic threat posed by new, emerging, reassortant influenza viruses, for which humans are immunologically naive, represents a major public health concern. Current influenza vaccines are impacted by several issues. These include, the elicitation of narrow, strain-specific immune responses, an over-reliance on egg-based manufacturing methods, a protracted production process (>6 months), the need to predict in advance which strains will circulate in forthcoming seasons and the minimal induction of cellular and humoral immune responses to multiple influenza antigens (Ags) simultaneously. The sub-optimal performance of seasonal influenza vaccines in recent years has accelerated interest in developing a universal influenza virus vaccine, capable of providing broad and long- lived protection against seasonal and pandemic subtypes. Strategies to achieve this include refocusing immune responses towards highly conserved epitopes on influenza virus antigens such as the stalk of the major surface glycoprotein, hemagglutinin (HA), the neuraminidase (NA) or the internal nucleoprotein (NP). My research aims to develop an alternative, optimized, universal influenza vaccine platform which will overcome issues associated with current vaccines using three approaches. (1) Firstly, I will optimize polycistronic Ag expression cassettes, in which multiple IAV Ags are expressed simultaneously. These will include bi- or tri-cistronic Ag cassettes featuring headless HAs from group 1 or group 2 IAVs in combination with NA and/or NP. I will augment/broaden immune recognition of headless HA or NA by targeting Ags to host- derived extracellular vesicles (EVs) including exosomes in vivo. This will be achieved by engineering fusion-Ag constructs to tether Ag to a protein domain enriched in exosomes. Exosomes are nano-sized EVs which play important roles in the regulation of immune responses, due to their ability to present Ag, in addition to MHC and co-stimulatory molecules, to T- and B-cells. (2) Secondly, I will engineer these Ag constructs into non- replicating, rare species adenoviral (Ad) vectored vaccines, which have established protocols for clinical manufacturing, can be thermostabilized with minimal losses to immunogenicity under cold-chain free conditions and have demonstrated safety and immunogenicity in infants, adults and the elderly in clinical trials. (3) Finally, I will comprehensively evaluate and phenotype the magnitude and profile of these universal influenza vaccines in single-shot regimens. These data will provide valuable information for the design of subsequent prime:boost regimens and for challenge experiments in the future. In summary, the universal influenza vaccine platform described in this proposal would be well-suited to stockpiling for pandemic preparedness, and could provide heterologous protection following a single shot, which may be sufficient to ease the burden on the healthcare system in the early phase of an emerging pandemic.

摘要：甲型流感病毒(IAV)可导致人类严重的呼吸道疾病，约有25万至50万 全球每年的死亡人数。除了季节性流行病外，新的、 新出现的、重新组合的流感病毒，人类在免疫学上是幼稚的，代表着一种主要的 公共卫生问题。目前的流感疫苗受到几个问题的影响。这些包括，启发式 狭隘的、菌株特异性的免疫反应，过度依赖以鸡蛋为基础的制造方法， 漫长的生产过程(&gt；6个月)，需要提前预测哪些菌株将在 即将到来的季节和对多重流感的细胞和体液免疫反应的最低限度诱导 抗原(AGS)同时存在。近几年季节性流感疫苗的次优表现 加速了开发通用流感病毒疫苗的兴趣，该疫苗能够提供广泛和长期的 针对季节性和大流行亚型的活保护。实现这一目标的战略包括调整重点 对流感病毒抗原高度保守表位的免疫应答 主要的表面糖蛋白、血凝素(HA)、神经氨酸酶(NA)或内部核蛋白(NP)。 我的研究旨在开发一个替代的、优化的、通用的流感疫苗平台，它将 使用三种方法克服与当前疫苗相关的问题。(1)首先，我会优化 多顺反子抗原表达盒，在其中同时表达多个IAV抗原。这些遗嘱 包括具有来自第1组或第2组IAV组合无头HAS的双顺反子或三顺反子银盒 带NA和/或NP。我将通过针对宿主的AGS来增强/扩大对无头HA或NA的免疫识别- 体内来源的细胞外小泡(EV)，包括外切体。这将通过工程融合-Ag来实现 构建了将Ag与富含外切体的蛋白质结构域连接的结构。Exosome是一种纳米大小的电动汽车，可以玩 在免疫反应调节中的重要作用，因为它们除了MHC外，还能呈递抗原 以及共刺激分子，转化为T细胞和B细胞。(2)其次，我将把这些Ag结构设计成非 复制的稀有种腺病毒(Ad)载体疫苗，已建立临床方案 制造，可以在无冷链的情况下以最小的免疫原性损失进行热稳定 在临床试验中，已证明对婴儿、成人和老年人具有安全性和免疫原性。 (3)最后，我将对这些普遍性的大小和轮廓进行全面的评估和表型 单针接种的流感疫苗。这些数据将为设计提供有价值的信息。 随后的主要内容：加强养生法，并在未来进行挑战实验。总而言之，普遍的 本提案中描述的流感疫苗平台将非常适合为大流行进行储备 防备，并可在单次射击后提供异种保护，这可能是 足以在新出现的大流行的早期阶段减轻医疗系统的负担。