Improved vaccine manufacture to control foot-and-mouth disease: Production of recombinant vaccines by design

改进疫苗生产以控制口蹄疫：通过设计生产重组疫苗

• 批准号: BB/N007298/1
• 负责人: Tobias Tuthill
• 金额: $ 44.93万
• 依托单位: The Pirbright Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN007298%2F1
• 关键词: Improved; vaccine; manufacture; control; foot

One of our biggest challenges will be to meet a growing demand for food especially in the developing world. Animal diseases have a significant impact on the productivity of the livestock industry and safeguarding animal welfare will be a major part of maximising food production. Foot-and-mouth disease (FMD) is endemic in many regions of the world and affects huge numbers of animals, both wild animals and livestock (e.g. cattle, sheep, goats, pigs) and is greatly feared due to the enormous economic losses associated with outbreaks. Thus improved control of FMD could sustainably improve the performance of livestock industries and have enormous economic and social value worldwide especially in developing regions where livestock are seen as a means to raise millions from poverty. In FMD-free regions, such as UK, the costs of FMD primarily result from the need to maintain a disease free status. However, FMD-free countries can face enormous losses as the result of periodic incursions and the estimated cost of the 2001 UK outbreak was £8-10Bn. Future incursions into the UK will undoubtedly occur with the potential to inflict substantial economic losses. Vaccination remains the most effective approach for controlling viral diseases and an estimated 2.35 billion doses of FMD vaccine are administered annually. Currently vaccination-led eradication campaigns are ongoing in India, South-East Asia and South America. Two major constraints on FMD vaccination are (i) production capacity and (ii) the need to produce new vaccines from field isolates. Production capacity: FMD vaccines are chemically-inactivated virus preparations produced in cell-culture and the highly contagious nature of FMD requires they are produced in expensive, high-containment facilities. Thus measure that improve vaccine yield from existing production facilities could have an enormous impact by increasing the amount of vaccine available globally for FMD control.New vaccines: FMD is caused by FMD virus (FMDV). A major problem for FMD control is the existence of seven FMDV serotypes, each formed by multiple and constantly evolving virus subtypes. Importantly, vaccination against one serotype does not protect against the others. Further, it is essential that within a serotype the vaccine offers protection against the circulating outbreak strain. This necessitates the periodic need to produce new vaccines, especially against emerging strains for which the current vaccines are a poor match. Vaccine production uses established cell lines (such as BHK cells) that are not normally targeted by FMDV. Thus, the production of new vaccines is critically dependent upon adaptation of a field virus for growth in cell-culture which can prove time consuming or impossible for some field viruses. Thus the challenge is to produce sufficient quantities of vaccine for FMD control We have shown that cell-culture adaptation of FMDV and improved cell-culture growth are achieved by specific mutations at either of two distinct sites in the viral capsid. It is our hypothesis that recombinant FMDV with such mutations will have in-built, improved cell-culture growth and can be used to overcome the delays in producing new vaccines from field viruses and, through improved cell-culture growth, increase vaccine yield from the existing manufacturing capacity. Here we will use reverse genetics to genetically engineer FMDV to include mutations in the viral capsid that result in improved virus growth and develop these viruses a vaccines. We will focus on four of the most prevalent serotypes and include viruses that are known to currently give a poor vaccine yield and have proven difficult to grow in cell-culture. Our ambition is to develop marketable products that could have enormous value by reducing the prevalence of FMD worldwide and the likelihood of further outbreaks of FMD in the UK and Europe.

我们面临的最大挑战之一将是满足日益增长的粮食需求，特别是在发展中国家。动物疾病对畜牧业的生产力有重大影响，保障动物福利将是最大限度地提高粮食产量的重要组成部分。口蹄疫（FMD）在世界许多地区是地方性的，并且影响大量动物，包括野生动物和家畜（例如牛、绵羊、山羊、猪），并且由于与爆发相关的巨大经济损失而非常令人担忧。因此，改善口蹄疫的控制可以可持续地改善畜牧业的绩效，并在世界范围内具有巨大的经济和社会价值，特别是在发展中地区，畜牧业被视为使数百万人脱贫的手段。在无口蹄疫地区，如英国，口蹄疫的成本主要来自于维持无病状态的需要。然而，由于周期性入侵，无口蹄疫国家可能面临巨大损失，2001年英国爆发的估计成本为80亿至100亿英镑。毫无疑问，未来对英国的入侵将有可能造成重大经济损失。疫苗接种仍然是控制病毒性疾病的最有效方法，估计每年接种23.5亿剂口蹄疫疫苗。目前，印度、东南亚和南美洲正在开展以接种疫苗为主导的根除运动。口蹄疫疫苗接种的两个主要限制因素是（i）生产能力和（ii）需要从田间分离株中生产新疫苗。生产能力：口蹄疫疫苗是在细胞培养中生产的化学灭活病毒制剂，口蹄疫的高度传染性要求它们在昂贵的高防护设施中生产。因此，提高现有生产设施疫苗产量的措施可能会产生巨大影响，增加全球可用于控制口蹄疫的疫苗数量。FMD控制的一个主要问题是存在七种FMDV血清型，每种血清型由多种和不断进化的病毒亚型形成。重要的是，针对一种血清型的疫苗接种不能保护其他血清型。此外，在血清型内，疫苗必须提供针对流行爆发毒株的保护。这就需要定期生产新疫苗，特别是针对目前疫苗不匹配的新菌株。疫苗生产使用通常不被FMDV靶向的已建立的细胞系（如BHK细胞）。因此，新疫苗的生产关键取决于田间病毒在细胞培养物中生长的适应性，这对于一些田间病毒来说可能是耗时的或不可能的。因此，挑战是生产足够量的疫苗用于FMD控制。我们已经表明，FMDV的细胞培养适应和改善的细胞培养生长是通过病毒衣壳中两个不同位点中任一个的特异性突变实现的。我们的假设是，具有这种突变的重组FMDV将具有内置的、改善的细胞培养生长，并且可以用于克服从田间病毒生产新疫苗的延迟，并且通过改善细胞培养生长，从现有的生产能力增加疫苗产量。在这里，我们将使用反向遗传学的基因工程口蹄疫病毒，包括在病毒衣壳的突变，导致改善病毒的生长和发展这些病毒的疫苗。我们将重点关注四种最流行的血清型，并包括已知目前疫苗产量较差且已证明难以在细胞培养中生长的病毒。我们的目标是开发适销对路的产品，通过减少全球口蹄疫的流行以及英国和欧洲进一步爆发口蹄疫的可能性，这些产品可能具有巨大的价值。

项目成果

Irreversible inactivation of ISG15 by a viral leader protease enables alternative infection detection strategies.

• DOI: 10.1073/pnas.1710617115
• 发表时间: 2018-03-06
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Swatek KN;Aumayr M;Pruneda JN;Visser LJ;Berryman S;Kueck AF;Geurink PP;Ovaa H;van Kuppeveld FJM;Tuthill TJ;Skern T;Komander D
• 通讯作者: Komander D

Generation and characterisation of recombinant FMDV antibodies: Applications for advancing diagnostic and laboratory assays.

• DOI: 10.1371/journal.pone.0201853
• 发表时间: 2018
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Shimmon G;Kotecha A;Ren J;Asfor AS;Newman J;Berryman S;Cottam EM;Gold S;Tuthill TJ;King DP;Brocchi E;King AMQ;Owens R;Fry EE;Stuart DI;Burman A;Jackson T
• 通讯作者: Jackson T