Developing a Paramyxovirus-based H5N1 Vaccine

开发基于副粘病毒的 H5N1 疫苗

• 批准号: 8134261
• 负责人: Biao He
• 金额: $ 70.96万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-25 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8134261
• 关键词: Affect; Age; Antigen-Presenting Cells; Antigens; Apoptosis; Apoptotic; Attenuated; Avian Influenza; Avian Influenza A Virus; Birds; Canis familiaris; Cell Death; Cells; Cessation of life; Clinical Trials; Coughing; Dendritic Cells; Disease Outbreaks; Distant; Dose; FDA approved; Gene Expression; Genes; Genetic Risk; Health; Hospitalization; Human; Human Cell Line; Immune response; Immunity; Immunization; Inactivated Vaccines; Individual; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A virus; Life; Morbidity - disease rate; Mus; National Institute of Allergy and Infectious Disease; Paramyxovirus; Population; Positioning Attribute; Principal Investigator; Production; Proteins; Recombinants; Reporting; Safety; Source; Southeastern Asia; Testing; Time; United States; Vaccinated; Vaccine Antigen; Vaccine Production; Vaccines; Vero Cells; Viral Antigens; Viral Proteins; Virus; Virus Diseases; age group; base; comparative efficacy; dosage; efficacy testing; flu; immunogenic; immunogenicity; improved; influenza virus gene; influenza virus strain; influenza virus vaccine; influenzavirus; interest; mortality; mutant; novel vaccines; pandemic disease; pandemic influenza; parainfluenza virus; positional cloning; prevent; programs; promoter; public health relevance; recombinant virus; vaccine candidate; vaccine development; vector; vector vaccine

DESCRIPTION (provided by applicant): H5N1 is a looming threat to human health. A safe and effective vaccine is the best way to prevent large-scale outbreaks in the human population. Parainfluenza virus 5 (PIV5), a paramyxovirus, causes kennel cough in dogs, but is not known to cause any illness in humans. PIV5 can be produced in high titers (up to 8x108 plaque forming units (PFU)/ml) in many cells including Vero cells, which are WHO approved for vaccine production. PIV5 can infect human cell lines as well as primary human cells. In our preliminary studies, a single dosage of 104 PFU of a live recombinant PIV5 expressing a HA gene from the H3 subtype provided immunity against influenza A virus subtype 3 infection in mice. We hypothesize that PIV5 is a good vector for vaccine development and we propose to test PIV5 as a vector for H5N1 vaccine. We will focus our efforts on following specific aims: 1. Testing recombinant PIV5 expressing HA of H5N1 (PIV5-H5) as a vaccine for H5N1; 2. Testing the potential of other H5N1 proteins expressed using PIV5 as antigens for vaccine; and 3. Testing mutant PIV5 viruses as vaccine vectors. PUBLIC HEALTH RELEVANCE: Influenza A virus causes significant morbidity and mortality each year. Strains currently circulating in humans (i.e. H1N1, H1N2, and H3N2) infect up to 15% of the world population and cause an average of 36,000 deaths and 226,000 hospitalizations in the United States (28), as well as millions deaths world wide. Sporadic outbreaks of pandemic influenza have caused significant mortality over the past century, most notably the Spanish flu of 1918, and have caused over 50 million deaths world wide. On the horizon is another potentially pandemic strain of influenza, H5N1. This avian influenza virus has most notably emerged in Southeast Asia and resulted in the destruction of millions of birds, infected 309 people, caused 187 human fatalities since 2003 (WHO, Cumulative Number of Confirmed Human Cases of Avian Influenza A/(H5N1) Reported to WHO, 31 May 2007), and threatens to become the next pandemic. Currently, the only FDA-approved vaccine against H5N1 has serious limitations, particularly as it has to be given twice and requires substantial higher concentrations of the vaccine to achieve a moderate level of efficacy compared to conventional influenza vaccines. Conventional vaccines utilizing the HA and NA of H5N1 viruses have been poorly immunogenic and have safety and production issues. A live-attenuated H5N1 vaccine has been generated by reverse genetics, but the risk of generating a reassortant prohibits use of this vaccine in most instances. Inactivated virus vaccines have also been derived by reverse genetics and produced in large quantities, but preliminary results from NIAID clinical trials suggest that efficacy will require both multiple immunizations and 6 times the standard influenza virus antigen dose, i.e. 90 ug instead of 15 ug of antigen while only providing protection in a subset (~50%) of vaccinated individuals. Nonetheless, FDA has recently approved the inactivated H5N1 vaccine for use in people between age 18 and 64, an age group that is not the most vulnerable to influenza virus infection. Thus, there is a rationale need for new vaccine strategies that provide increased immunogenicity and safety. Parainfluenza virus 5 (PIV5), a paramyxovirus, causes kennel cough in dogs, but is not known to cause any illness in humans. PIV5 can be produced in high titers (up to 8x108 plaque forming units (PFU)/ml) in many cells including Vero cells, which are WHO-approved for vaccine production. PIV5 can infect human cell lines as well as primary human cells. In our preliminary studies, a single dosage of 104 PFU of a live recombinant PIV5 expressing a HA gene from the H3 subtype provided immunity against influenza A virus subtype 3 infection in mice. In this proposal, we hypothesize that PIV5 is a good vector for vaccine development and we propose to test PIV5 as a vector for H5N1 vaccine.

描述（由申请人提供）：H5N1是对人类健康的迫在眉睫的威胁。安全有效的疫苗是防止人口大规模爆发的最佳方法。 Parainfluenza病毒5（PIV5），一种帕托马病毒，会引起狗的狗窝咳嗽，但尚不知道会在人类中引起任何疾病。 PIV5可以在包括Vero细胞在内的许多细胞中以高滴度（高达8x108斑块形成单元（PFU）/mL）生产，这些细胞已批准用于疫苗的生产。 PIV5可以感染人类细胞系以及原代人细胞。在我们的初步研究中，从H3亚型中表达HA基因的活体重组PIV5的单个剂量为小鼠中的流感A亚型3感染提供了免疫力。我们假设PIV5是疫苗开发的良好载体，我们建议将PIV5作为H5N1疫苗的载体进行测试。我们将重点放在遵循特定目标上：1。测试重组PIV5的H5N1（PIV5-H5）作为H5N1的疫苗； 2。测试使用PIV5作为疫苗抗原表达的其他H5N1蛋白的潜力； 3。将突变PIV5病毒作为疫苗载体。 公共卫生相关性：每年流感病毒会引起大量发病率和死亡率。目前在人类中流通的菌株（即H1N1，H1N2和H3N2）感染了多达15％的世界人口，并在美国平均导致36,000人死亡和226,000例住院（28），以及全球数百万的死亡人数。大流行性流感的零星爆发在过去一个世纪中引起了重大死亡率，最著名的是1918年的西班牙流感，并且在全球造成了超过5000万人死亡。地平线是流感的另一种潜在大流行菌株H5N1。这种鸟类流感病毒最著名的是在东南亚出现，并导致数百万只鸟类被感染，感染了309人，自2003年以来造成了187人死亡（谁，累积的确认人类禽流感A/（H5N1）报告给WHO，2007年5月31日向WHO报告，并威胁到下一个PARDEMENT。 当前，唯一针对H5N1的FDA批准的疫苗具有严重的局限性，尤其是必须给出两次，并且与常规的流感疫苗相比，它需要更高浓度的疫苗才能达到中等水平的疗效。利用H5N1病毒的HA和NA的常规疫苗的免疫原性很差，并且存在安全性和生产问题。反向遗传学产生了活体衰减的H5N1疫苗，但是在大多数情况下，可能会产生重新成分的疫苗的风险。灭活的病毒疫苗也是通过反向遗传学得出的，并大量产生，但是NIAID临床试验的初步结果表明，功效将需要多次免疫和6倍的标准流感病毒抗原剂量的6倍，即90 ug，而不是仅提供15 ug antigen and Antigen，而在抗原中只能提供替代（〜50％）的抗量（〜50％）。尽管如此，FDA最近批准了18至64岁的人使用的灭活H5N1疫苗，该年龄段并非最容易受到流感病毒感染的影响。因此，需要新的疫苗策略的理由，这些疫苗策略提供了提高的免疫原性和安全性。 Parainfluenza病毒5（PIV5），一种帕托马病毒，会引起狗的狗窝咳嗽，但尚不知道会在人类中引起任何疾病。 PIV5可以在许多细胞（包括Vero细胞）中以高滴度（高达8x108斑块形成单元（PFU）/mL）生产，WHO批准了用于疫苗的生产。 PIV5可以感染人类细胞系以及原代人细胞。在我们的初步研究中，从H3亚型中表达HA基因的活体重组PIV5的单个剂量为小鼠中的流感A亚型3感染提供了免疫力。在此提案中，我们假设PIV5是疫苗开发的良好载体，我们建议将PIV5作为H5N1疫苗的载体进行测试。