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.

工程化鼻内通用流感疫苗 流感病毒引起严重的呼吸道疾病。由于流感基因的高突变率， 漂流每年都会产生一种新的菌株因此，监测病毒存在重大的经济负担 活动，并创造和分发新的流感疫苗给公众每年。此外，由于基因 重组，一种新的流感亚型可能会出现，这是致命的，从来没有流传之间 人类，并可能导致毁灭性的流行病。因此，至关重要的是，与当前的疫苗不同， 制定针对甲型流感的通用疫苗接种策略，以保护免受所有甲型流感的侵害 亚型病毒跨膜蛋白M2（M2 e）的23个氨基酸长的胞外结构域发现于 自1918年流感大流行以来，人（h）甲型流感病毒一直保持高度保守;因此，hM 2 e是 被认为是开发通用甲型流感疫苗的良好候选者。然而，hM 2 e是差的， 免疫原性。我们设计了hM 2 e的鼻内给药系统。我们的研究表明， hM 2 e与病毒模拟金纳米颗粒（AuNPs）的结合以及使用CpG作为可溶性佐剂（AuNP-hM 2 e）， hM 2 e +sCpG疫苗），在Balb/c小鼠中可观察到针对人H1N1的广泛异亚型保护 和H3 N2甲型流感毒株，以及高致病性禽流感毒株H5 N1。然而，共识 人（h）、禽（a）和猪（s）流感病毒中M2 e的氨基酸序列显示出变异性。确实 我们的初步研究表明，鼻内接种AuNP-hM 2 e +sCpG疫苗的小鼠仅 对安徽/1/2013（H7N9）的致命攻击有部分保护，这是一种禽流感病毒，和 其M2 e序列与禽类aM 2 e的同源性高于hM 2 e。令人惊讶的是，我们的疫苗也可以诱导 C57 BL/6小鼠和CD-1小鼠（远交品系）中的M2 e抗体应答，从而证明了 适用于遗传多样性的人群。因此，我们假设，基于 AuNP-M2 e +sCpG并且包含hM 2 e、aM 2 e和sM 2 e共有序列作为抗原的抗体可以是 开发了一种广泛的交叉保护，持久和安全的甲型流感疫苗， 基因多样的人群。我们的具体目标是：（一）建立遗传学交叉保护的广度 不同的小鼠品系，并评估长期疫苗保护。(ii)确定免疫机制 疫苗的刺激。(iii)确定疫苗的生物分布和安全性。 如果成功的话，这些研究将提供必要的有效性和安全性数据，以支持疫苗的推广。 到第一阶段临床试验