Novel Bioconjugate Vaccines to Prevent Staphylococcus aureus Infection

预防金黄色葡萄球菌感染的新型生物结合疫苗

基本信息

批准号：
8637906
负责人：
Jean Claire Lee
金额：
$ 66.94万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8637906
关键词：
Abbreviations Active Immunization Anabolism Animal Model Animals Antibiotics Antibodies B-Lymphocyte Epitopes Bacteremia Biotechnology Bovine Serum Albumin Campylobacter Carrier Proteins Catheters Cells Communities Complex Conjugate Vaccines Consensus Sequence Contracts Coupled Data Development Disease Encapsulated Endocarditis Engineering Enzymes Escherichia coli Foundations Glycoproteins Goals Human Immune Immune Sera Infection Intravenous Life Link Methicillin Resistance Modeling Mus Oligosaccharides Oryctolagus cuniculus Panton-Valentine leukocidin Passive Immunization Peptides Pharmacologic Substance Pneumonia Polysaccharides Process Production Protein Glycosylation Proteins Pseudomonas aeruginosa Rattus Recombinants Rodent Model Serum Staphylococcal Infections Staphylococcus aureus Streptomycin Structure Surgical Wound Infection Switzerland System T-Cell Depletion T-Lymphocyte Technology Testing Toxoids Vaccines Wound Infection alpha Toxin base capsule cost effective efficacy testing factor A immunogenicity in vitro Assay intraperitoneal killings methicillin resistant Staphylococcus aureus microbial alkaline proteinase inhibitor neutrophil new technology novel novel vaccines pathogen periplasm prevent research study safety testing subcutaneous undecaprenyl pyrophosphate

项目摘要

DESCRIPTION (provided by applicant): The overall objective of this project is the development of an effective vaccine to prevent invasive, life- threatening infections by the medically important bacterial pathogen Staphylococcus aureus. Because S. aureus cannot always be controlled by antibiotics and methicillin-resistant isolates are becoming increasingly prevalent in the community, a S. aureus vaccine is sorely needed. GlycoVaxyn, a small biotechnology firm in Switzerland, has developed a proprietary technology that enables the manufacture of bioconjugate vaccines based on complex polysaccharide structures. A novel Campylobacter enzyme called PglB is able to transfer an oligosaccharide to a protein consensus sequence, thereby allowing the production of glycoproteins in bacterial cells. This protein glycosylation system has been functionally transferred into Escherichia coli, and it allows the conjugation of an antigenic polysaccharide to a designer protein of choice through an N-glycosidic linkage. Conjugated vaccines have been produced in E. coli by co-expression of PglB, a protein carrier, and an antigenic polysaccharide cluster. In this approach, the conjugated vaccine can be extracted from the periplasm of E. coli and purified. In preliminary studies GlycoVaxyn has prepared a novel vaccine by engineering E. coli to express the S. aureus capsule type 5 polysaccharide (CP5) vaccine covalently linked to the carrier protein EPA (nontoxic Pseudomonas aeruginosa exoprotein A). The vaccine was purified, characterized, and injected into mice and rabbits to demonstrate its immunogenicity. High titered rabbit antibodies to CP5-EPA promoted opsonophagocytic killing of two different S. aureus strains in an in vitro assay with human neutrophils. In addition, passive transfer of CP5-EPA antibodies resulted in a 98% reduction in S. aureus bacteremia in mice. These data provide a strong foundation for the proposed studies to determine whether multicomponent S. aureus bioconjugate vaccines will protect against disease provoked by multiple S. aureus strains in a variety of infection models. Our specific aims are to (1) utilize GlycoVaxyn's proprietary glyco-engineering technology to produce a tetravalent bioconjugate vaccine against S. aureus: CP5-clumping factor A (ClfA) and CP8-alpha toxoid; (2) perform in vitro assays to evaluate the functionality of antibodies elicited by the bioconjugate vaccines; (3) perform active immunization experiments to test the efficacy of the multi-component bioconjugate vaccine in rodent models of staphylococcal infection, including bacteremia, pneumonia, wound infection, and endocarditis; (4) investigate the immune correlates of protection against S. aureus infection by passive immunization and T cell depletion experiments; (5) development of a GMP compatible production process in E. coli for the tetravalent bioconjugate. By glyco-engineering technology, capsular polysaccharides are conjugated to known protective peptides without denaturing the protein carrier, thus conserving B cell epitopes. This approach paves the way for the biosynthesis of previously unattainable vaccine materials in a cost effective manner. The overall goal of this project is the development of an effective vaccine to prevent invasive, life-threatening infections by the medically important bacterial pathogen Staphylococcus aureus. A new technology allows for bacterial synthesis of vaccine proteins coupled to polysaccharides with known vaccine potential. The "bioconjugate" vaccines will be tested in animal models of staphylococcal infection and processed for initial safety testing in humans.

描述（由申请人提供）：该项目的总体目的是开发有效的疫苗，以防止医学上重要的细菌病原体葡萄球菌金黄色葡萄球菌的侵入性，威胁生命的感染。由于金黄色葡萄球菌不能总是由抗生素控制，耐甲氧西林的分离株在社区中变得越来越普遍，因此迫切需要一种金黄色葡萄球菌疫苗。瑞士的一家小型生物技术公司Glycovaxyn开发了一种专有技术，该技术可以基于复杂的多糖结构来生产生物偶联疫苗。一种称为PGLB的新型弯曲杆菌能够将寡糖转移到蛋白质共识序列上，从而允许在细菌细胞中产生糖蛋白。该蛋白质糖基化系统已在功能上转移到大肠杆菌中，并允许通过N-糖苷键将抗原性多糖偶联到选择的蛋白质。通过蛋白质载体PGLB和抗原多糖簇的PGLB共表达，已在大肠杆菌中产生了共轭疫苗。在这种方法中，可以从大肠杆菌的周期中提取共轭疫苗并纯化。在初步研究中，Glycovaxyn通过工程大肠杆菌制备了一种新型的疫苗，以表达与载体蛋白EPA（无毒假单胞菌铜绿氧化物甲基糖蛋白A）的载体蛋白EPA相关的5多糖（CP5）疫苗。将疫苗纯化，表征并注入小鼠和兔子，以证明其免疫原性。对CP5-EPA的高滴度兔抗体促进了与人类嗜中性粒细胞的体外测定中两种不同的金黄色葡萄球菌菌株的op子杀伤。此外，CP5-EPA抗体的被动转移导致小鼠金黄色葡萄球菌的降低98％。这些数据为拟议的研究提供了一个巨大的基础，以确定多组分金黄色葡萄球菌生物缀合物疫苗是否会预防多种感染模型中多个金黄色葡萄球菌菌株引起的疾病。我们的具体目的是（1）利用甘氨酸的专有的Glyco工程技术来生产针对金黄色葡萄球菌的四位价值生物偶联物疫苗：CP5倾斜因子A（CLFA）和CP8-Alpha-Alpha Toxoid; （2）进行体外测定，以评估生物偶联物疫苗引起的抗体的功能；（3）进行主动免疫实验，以测试多组分生物缀合物疫苗在葡萄球菌感染的啮齿动物模型中的功效，包括菌血症，肺炎，伤口感染和心内膜炎；（4）通过被动免疫和T细胞耗竭实验研究保护对金黄色葡萄球菌感染的免疫相关性；（5）在四二元生物缀合物的大肠杆菌中开发GMP兼容的生产过程。通过Glyco工程技术，将囊囊多糖与已知的保护性肽偶联而不贬低蛋白质载体，从而保存B细胞表位。这种方法为以前无法实现的疫苗材料的生物合成铺平了道路。该项目的总体目标是开发有效的疫苗，以防止医学上重要的细菌病原体葡萄球菌金黄色葡萄球菌的侵入性，威胁生命的感染。一种新技术允许细菌合成与已知疫苗潜力的多糖结合的疫苗蛋白。将在葡萄球菌感染的动物模型中测试“生物轭”疫苗，并在人类中进行初步的安全测试。