Development and testing of novel recombinant pnemococcal glyconjugate vaccines

新型重组肺炎球菌糖复合物疫苗的开发和测试

• 批准号: MR/K012053/1
• 负责人: Brendan Wren
• 金额: $ 71.96万
• 依托单位: London School of Hygiene & Tropical Medicine
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK012053%2F1
• 关键词: Development; testing; novel; recombinant; pnemococcal

Streptococcus pneumoniae, or the pneumococcus, can cause life-threatening diseases such as pneumonia, septicaemia, meningitis and frequently causes ear infections in children which can lead to hearing loss. S. pneumoniae is responsible for significant morbidity and mortality worldwide and by conservative estimates pneumococcal infections cause over one million deaths of children annually. An inexpensive, broad-range, long-lasting pneumococcal vaccine is desperately required.A defining characteristic of a successful vaccine is the ability to evoke long-lasting protective immunity with minimal side effects. The most successful human vaccines are often glycoconjugate as the combination of a protein coupled to a sugar glycan induces both a T-cell dependent and independent immune response evoking a protective and lasting immunity. Examples of currently licensed human glycoconjugate vaccines include those against Haemophilus influenzae, Neisserria meningitidis and some Streptococcus pneumoniae strains, in which glycans are chemically coupled to immunogenic carrier proteins.Traditional glycoconjugate vaccine design by chemical conjugation requires that the glycan from the pathogenic organism be isolated, detoxified by stripping out surface components, and still be present in sufficient amounts to be chemically coupled to a protein. The procedures involve harsh chemical treatments, are time consuming and expensive. In addition, the material generated at each step needs to be verified for purity, and variation between batches of glycoconjugate vaccine is common. Current licensed pneumococcus glycoconjugate vaccines are problematic as they only cover a fraction of all S. pneumoniae strains. Although there are vaccine candidates based on conserved proteins, these vaccines often do not produce long-term protection that is especially required to immunize the main target population, children. Ideally, a glycoconjugate vaccine based on conserved pneumococcal proteins coupled to the capsular polysaccharide glycan should be produced, but to date this has proved technically challenging.Recently, we (and collaborators) have developed a new approach for constructing glycoconjugate vaccines involving cloning all components in the widely used "work-horse" microbe E. coli. The recombinant process is termed Protein Glycan Coupling Technology (PGCT) and involves processing the candidate protein and glycan in plasmid vectors in E. coli along with a coupling enzyme to produce an inexhaustible supply of vaccine. PGCT can produce purified vaccine in a one-step purification procedure, which reduces costs, and because multiple combinations of protein and glycans can be coupled together, a greater flexibility in the range of vaccines can be generated and tested. We will use PGCT to produce and test six outstanding protein candidates coupled to different combinations of pneumococcal capsular polysaccharide. These vaccines will be tested in the murine pneumococcal infection model for their relative protection against an otherwise lethal dose. Additionally, the vaccines will be tested for their effect on the carriage of pneumococci in the murine model. The new vaccines generated in this study will also be compared to the efficacy of existing pneumococcal vaccines such as Prevnar13. Data between experiments will be evaluated to derive the most efficacious glycoconjugate vaccine combination produced by PGCT. Additionally, the development of PGCT in this study will provide the expertise and knowledge base to make the technology more widely applicable to construct further S. pneumoniae glycoconjugate vaccines and vaccines against other important infectious agents.

肺炎链球菌，或肺炎球菌，可导致危及生命的疾病，如肺炎，败血症，脑膜炎，并经常导致儿童耳部感染，可导致听力损失。S.肺炎是世界范围内发病率和死亡率较高的原因，保守估计，肺炎球菌感染每年导致超过100万儿童死亡。目前迫切需要一种廉价、广谱、持久的肺炎球菌疫苗。成功疫苗的一个明确特征是能够激发持久的保护性免疫力，副作用最小。最成功的人类疫苗通常是糖缀合物，因为与糖聚糖偶联的蛋白质的组合诱导T细胞依赖性和非依赖性免疫应答，从而引起保护性和持久的免疫。目前许可的人糖缀合物疫苗的实例包括针对流感嗜血杆菌、脑膜炎奈瑟氏球菌和一些肺炎链球菌菌株的那些，其中聚糖与免疫原性载体蛋白化学偶联。并且仍然以足以与蛋白质化学偶联的量存在。这些程序涉及苛刻的化学处理，既耗时又昂贵。此外，每个步骤生成的材料需要验证纯度，并且糖缀合物疫苗批次之间的变化是常见的。目前许可的肺炎球菌糖缀合物疫苗是有问题的，因为它们仅覆盖所有S的一小部分。pneumoniae菌株虽然有基于保守蛋白的候选疫苗，但这些疫苗通常不能产生对主要目标人群（儿童）免疫所特别需要的长期保护。理想情况下，应该生产基于保守的肺炎球菌蛋白偶联到荚膜多糖聚糖的糖缀合物疫苗，但迄今为止，这已被证明在技术上具有挑战性。最近，我们（和合作者）已经开发出一种新的方法，用于构建糖缀合物疫苗，包括克隆广泛使用的“工作马”微生物E.杆菌重组过程称为蛋白聚糖偶联技术（PGCT），包括在大肠杆菌中处理质粒载体中的候选蛋白和聚糖。大肠杆菌沿着一种偶联酶来生产取之不尽的疫苗。PGCT可以在一步纯化程序中生产纯化的疫苗，这降低了成本，并且由于蛋白质和聚糖的多种组合可以偶联在一起，因此可以产生和测试疫苗范围的更大灵活性。我们将使用PGCT来生产和测试六种与肺炎球菌荚膜多糖不同组合偶联的优秀蛋白候选物。这些疫苗将在鼠肺炎球菌感染模型中进行测试，以确定其对其他致死剂量的相对保护作用。此外，将在鼠模型中检测疫苗对肺炎球菌携带的影响。本研究中产生的新疫苗也将与现有肺炎球菌疫苗（如Prevnar13）的有效性进行比较。将评价实验之间的数据，以得出PGCT生产的最有效的糖缀合物疫苗组合。此外，本研究中PGCT的发展将提供专业知识和知识基础，使该技术更广泛地适用于构建进一步的S。肺炎糖缀合物疫苗和针对其它重要传染因子的疫苗。

项目成果

Comparative Genomic Analysis and In Vivo Modeling of Streptococcus pneumoniae ST3081 and ST618 Isolates Reveal Key Genetic and Phenotypic Differences Contributing to Clonal Replacement of Serotype 1 in The Gambia.

• DOI: 10.1093/infdis/jix472
• 发表时间: 2017-12-05
• 期刊: The Journal of infectious diseases
• 作者: Bricio-Moreno L;Ebruke C;Chaguza C;Cornick J;Kwambana-Adams B;Yang M;Mackenzie G;Wren BW;Everett D;Antonio M;Kadioglu A
• 通讯作者: Kadioglu A

Population biology of Streptococcus pneumoniae in West Africa: multilocus sequence typing of serotypes that exhibit different predisposition to invasive disease and carriage.

• DOI: 10.1371/journal.pone.0053925
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Donkor ES;Adegbola RA;Wren BW;Antonio M
• 通讯作者: Antonio M

Temporal changes in nasopharyngeal carriage of Streptococcus pneumoniae serotype 1 genotypes in healthy Gambians before and after the 7-valent pneumococcal conjugate vaccine.

• DOI: 10.7717/peerj.903
• 发表时间: 2015
• 期刊: PeerJ
• 影响因子: 2.7
• 作者: Ebruke C;Roca A;Egere U;Darboe O;Hill PC;Greenwood B;Wren BW;Adegbola RA;Antonio M
• 通讯作者: Antonio M

Production and efficacy of a low-cost recombinant pneumococcal protein polysaccharide conjugate vaccine.

• DOI: 10.1016/j.vaccine.2018.05.036
• 发表时间: 2018-06-18
• 期刊: Vaccine
• 影响因子: 5.5
• 作者: Herbert JA;Kay EJ;Faustini SE;Richter A;Abouelhadid S;Cuccui J;Wren B;Mitchell TJ
• 通讯作者: Mitchell TJ

Construction of a pneumolysin deficient mutant in streptococcus pneumoniae serotype 1 strain 519/43 and phenotypic characterisation.

肺炎链球菌血清型 1 菌株 519/43 中肺炎球菌溶血素缺陷突变体的构建和表型表征。

• DOI: 10.1016/j.micpath.2020.103999
• 发表时间: 2020
• 期刊: Microbial pathogenesis
• 影响因子: 3.8
• 作者: Terra VS