Engineering an intranasal universal influenza vaccine

设计鼻内通用流感疫苗

• 批准号: 9526306
• 负责人: Harvinder Singh Gill
• 金额: $ 58.43万
• 依托单位: TEXAS TECH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-09 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9526306
• 关键词: Adjuvant; Agonist; Amino Acid Sequence; Amino Acids; Antibody Response; Antigens; Avian Influenza; Avian Influenza A Virus; B-Lymphocytes; Binding; Biodistribution; Birds; C57BL/6 Mouse; California; Cells; Child; Consensus; Consensus Sequence; Data; Development; Dose; Economic Burden; Elderly; Electron Microscopy; Engineering; Epitopes; Exhibits; Extracellular Domain; Family suidae; Freeze Drying; Genes; Gills; Goals; Gold; Grant; Haemophilus influenzae; Haplotypes; Histocompatibility; Histopathology; Human; IgG2; Immobilization; Immune; Immune response; Immunization; Immunoglobulin G; Inbred BALB C Mice; Inbreeding; Inflammation; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Integral Membrane Protein; Investigation; Kidney; Knock-out; Liver; Longevity; M2 protein; Measures; Monitor; Morbidity - disease rate; Mouse Strains; Mus; Mutation; Nasal Epithelium; National Institute of Allergy and Infectious Disease; Pathogenicity; Pattern; Phase I Clinical Trials; Population Heterogeneity; Porcine Influenza A Virus; Protein Isoforms; Reassortant Viruses; Safety; Serum; Signal Transduction; Single-Stranded DNA; System; T-Lymphocyte; T-Lymphocyte Epitopes; TLR9 gene; Testing; Tissues; Transgenic Organisms; Vaccinated; Vaccination; Vaccine Design; Vaccines; Vietnam; Viral; Virulent; Virus; Zoonoses; base; circulating biomarkers; design; genetic makeup; immunogenic; immunogenicity; influenza virus vaccine; influenzavirus; lymph nodes; mouse model; nanoparticle; novel; pandemic disease; pandemic influenza; respiratory; response; swine influenza; uptake; vaccination strategy; vaccine safety

Engineering an intranasal universal influenza vaccine Influenza virus causes serious respiratory illness. Due to the high mutation rate in influenza genes, antigenic drift can create a new strain each year. Consequently there is significant economic burden to monitor virus activity and to create and distribute new influenza vaccines to the public each year. Furthermore, due to gene reassortment, a novel influenza subtype could emerge, which is virulent and has never circulated amongst humans, and could cause a devastating pandemic. It is therefore crucial that unlike current vaccines, more universal vaccination strategies be developed against influenza A that can protect against all influenza A subtypes. The 23 amino acid-long extracellular domain of the viral transmembrane protein M2 (M2e) found on human (h) influenza A viruses has remained highly conserved since the 1918 pandemic; and hM2e is thus considered a good candidate for the development of a universal influenza A vaccine. However, hM2e is poorly immunogenic. We have designed an intranasal delivery system for hM2e. Our studies show that by attaching hM2e to virus-mimicking gold nanoparticles (AuNPs) and by using CpG as a soluble adjuvant (AuNP- hM2e+sCpG vaccine) a broad heterosubtypic protection can be observed in Balb/c mice against human H1N1 and H3N2 influenza A strains, and the highly pathogenic avian influenza strain H5N1. However, the consensus amino acid sequence of M2e in human (h), avian (a), and swine (s) influenza viruses shows variability. Indeed our preliminary study shows that mice vaccinated intranasally by the AuNP-hM2e+sCpG vaccine are only partially protected against a lethal challenge by Anhui/1/2013 (H7N9), which is an avian reassortant virus, and whose M2e sequence has more homology to avian aM2e than hM2e. Amazingly, our vaccine can also induce M2e antibody response in C57BL/6 mice and CD-1 mice (an outbred strain), thus demonstrating an ability to be applicable to a genetically diverse population. We thus hypothesize that a multivalent vaccine based on AuNP—M2e+sCpG and comprising of hM2e, aM2e, and sM2e consensus sequences as antigens can be developed in to a broadly cross-protective, durable and safe influenza A vaccine that is effective in a genetically diverse population. Our specific aims are: (i) Establish breadth of cross-protection in genetically diverse mouse strains and evaluate long term vaccine protection. (ii) Determine mechanism of immune stimulation by the vaccine. (iii) Determine biodistribution and safety of the vaccine. If successful these studies will provide the efficacy and safety data necessary to support moving the vaccine in to phase-I clinical trials.

研制鼻腔通用流感疫苗 流感病毒会引起严重的呼吸道疾病。由于流感基因的高突变率，抗原性 漂移每年都会产生新的菌株。因此，监测病毒会带来巨大的经济负担。 每年开发并向公众分发新的流感疫苗。此外，由于基因 重新分类，可能会出现一种新的流感亚型，这种亚型是致命的，从未在 人类，并可能导致毁灭性的大流行。因此，与目前的疫苗不同的是，更多的疫苗 制定针对甲型流感的通用疫苗接种战略，以预防所有甲型流感 子类型。发现的病毒跨膜蛋白M2(M2e)的23个氨基酸的胞外区 人类(H)甲型流感病毒自1918年大流行以来一直保持高度保守；因此hM2e 被认为是开发通用甲型流感疫苗的良好候选者。然而，hM2e的表现很差。 免疫原性。我们设计了一种hM2e鼻腔给药系统。我们的研究表明，通过依恋 HM2e转化为模拟病毒的金纳米颗粒(AuNPs)，并将CpG作为可溶性佐剂(AuNP- HM2e+sCpG疫苗)可在Balb/c小鼠中观察到抗人H1N1流感的广泛异型保护作用 和H3N2型甲型流感病毒株，以及高致病性禽流感病毒株H5N1。然而，人们的共识是 人(H)、禽(A)和猪(S)流感病毒M2e的氨基酸序列具有变异性。确实如此 我们的初步研究表明，经鼻接种AuNP-hM2e+sCpG疫苗的小鼠仅 部分抵御安徽/1/2013(H7N9)的致命性挑战，这是一种禽类重组病毒，以及 其M2e序列与禽类aM2e的同源性高于hM2e。令人惊讶的是，我们的疫苗还可以诱导 在C57BL/6小鼠和CD-1小鼠(近交系)中的M2E抗体反应，从而表明有能力 适用于遗传多样性的人群。因此，我们假设一种多价疫苗基于 AUNP-M2e+sCpG和由hM2e、aM2e和sM2e共同序列组成的抗原可以是 发展成为广泛交叉保护、持久和安全的甲型流感疫苗，对 遗传多样性的种群。我们的具体目标是：(一)在遗传方面建立广泛的交叉保护 不同的小鼠品系，并评估长期疫苗保护。(二)确定免疫机制 疫苗的刺激作用。(3)确定疫苗的生物分布和安全性。 如果成功，这些研究将提供必要的有效性和安全性数据，以支持疫苗的进入 一期临床试验。