In vivo conjugated multivalent toxoid-polysaccharide vaccine for S. aureus

金黄色葡萄球菌体内多价类毒素-多糖缀合疫苗

• 批准号: 8645454
• 负责人: Rajan P Adhikari
• 金额: $ 30万
• 依托单位: INTEGRATED BIOTHERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8645454
• 关键词: Abscess; Adjuvant; Adult; Advanced Development; Agreement; Amino Acids; Anabolism; Animal Model; Antibiotics; Antibodies; Antigens; Attenuated; Bacteremia; Bacteria; Bacterial Infections; Biological Assay; Biological Response Modifier Therapy; Blood; Campylobacter; Cell Surface Proteins; Cells; Cessation of life; Chemicals; Chimeric Proteins; Chromatography; Clinical; Combined Vaccines; Conjugate Vaccines; Consensus Sequence; Data; Development; Disease; Disease model; Distant; Enterotoxins; Enzymes; Erythrocytes; Escherichia coli; Evaluation; Future; Genes; Glycoconjugates; Goals; Grant; Growth; Health; Hemolysin; Hospitalization; Human; Hybrids; Immune response; Individual; Infection; Intellectual Property; Joints; Kidney; Lead; Legal patent; Leucocidin; Leukocytes; Life; Link; Lipids; Liver; Lung; Lytic; Mass Spectrum Analysis; Measurement; Modeling; Monosaccharides; Mus; Mutation; National Institute of Allergy and Infectious Disease; Organ; Pathogenicity; Pathology; Patients; Peptide Signal Sequences; Peptides; Phase; Phase I Clinical Trials; Phenols; Pneumonia; Polysaccharides; Positioning Attribute; Preclinical Testing; Preventive; Production; Protein Glycosylation; Protein translocation; Proteins; Research; Rights; Risk; Safety; Sepsis; Serological; Shigella Vaccines; Site; Skin; Skin Tissue; Small Business Innovation Research Grant; Soft Tissue Infections; Solubility; Spleen; Staging; Staphylococcus aureus; Superantigens; Technology; Testing; Tissues; Toxin; Toxoids; Translational Research; United States; Vaccines; Virulence Factors; Western Blotting; base; biophysical properties; cellular engineering; design; dolichyl-diphosphooligosaccharide - protein glycotransferase; experience; flexibility; glycosylation; immunogenicity; in vivo; methicillin resistant Staphylococcus aureus; mouse model; neutrophil; new technology; novel; pathogen; periplasm; prevent; programs; protective efficacy; protein expression; public health relevance; resistant strain; screening; skin lesion; subcutaneous; vaccine candidate

DESCRIPTION (provided by applicant): Staphylococcus aureus is a Gram-positive human pathogen that causes a wide range of infections from skin and soft tissue infections (SSTI) to life threatening sepsis and pneumonia. The pathogenicity of S. aureus is dependent on numerous virulence factors, including cell surface proteins and polysaccharides, as well as secreted toxins. An important group of these toxins includes pore forming toxins such as hemolysins and leukocidins, as well as small peptidic toxins, with lytic activity towards erythrocytes and leukocytes particularly the neutrophils that represent the major line of defense against S. aureus. These toxins cause tissue damage, promote bacterial dissemination and growth in distant organs, and enable the pathogen to evade the host innate immune response. Furthermore, capsular polysaccharide (CP) are known to protect bacteria from phagocytic activity of polymorphonuclear leukocytes and have been validated as important vaccine targets. Integrated BioTherapeutics (IBT) is pursuing development of toxin- based vaccines in combination with CPs against S. aureus infections. The pore-forming ?-hemolysin (Hla), also known as ?-toxin (AT), is produced by nearly all strains and is implicated in several S. aureus invasive diseases. Recent microbiological and serological studies in humans with S. aureus bacteremia also show the importance of ¿-toxin and phenol-soluble modulins (PSM) for S. aureus pathogenicity. Based on our recent study, pre-existing antibodies against Hla, ¿-toxin and PSM?3 significantly reduce the risk of sepsis in adults with S. aureus bacteremia. The goal of this Phase I SBIR is to evaluate the feasibility of a novel glycoconjugate staphylococcal toxin based vaccine. Vaccine candidates have been designed that represent a critical structural domain at the N terminus of Hla (AT62) fused to ¿-toxin and PSM?3. We have shown previously that AT62 vaccine provides protection against bacteremia and pneumonia. This proposal utilizes a unique and novel in vivo bioconjugation technology developed by Glycovaxyn Inc. to conjugate this hybrid toxoid with capsular polysaccharide Type 5 (CP5) as proof of concept of the efficacy of the bioconjugate. The bioconjugation technology involves E. coli cells engineered to express the Campylobacter enzyme PglB as well as the genes required for S. aureus CP8 biosynthesis. PglB transfers N-linked CP8 chains to the vaccine construct introduced into these cells. This novel technology is a major development in glycoconjugate vaccine field with multiple advantages over the conventional chemical conjugation. In the Specific Aim 1 fusions of the three toxins will be generated and attenuating mutations identified and incorporated into the fusion construct. The candidate vaccine will be tested for immunogenicity and then bioconjugates generated in Aim 2. In Aim 3 the protective efficacy of the vaccine candidate will be evaluated in two major disease models representing bacteremia/sepsis and skin and soft tissue infections. Upon successful completion of this proposed research we envision a Phase II SBIR in which similar bioconjugate will be produced for the other major S. aureus capsular polysaccharide CP8 and the efficacy of the multivalent vaccine (including combination other toxoid vaccines currently under development in our group) extensively tested. The ultimate goal is to build a strategic partnership with large pharma to transition the vaccine candidate into clinical development.

描述（由适用提供）：金黄色葡萄球菌是一种革兰氏阳性的人类病原体，会引起从皮肤和软组织感染（SSTI）到威胁脓毒症和肺炎的生命的广泛感染。金黄色葡萄球菌的致病性取决于许多病毒因素，包括细胞表面蛋白和多糖以及分泌的毒素。这些毒素的重要组包括形成毒素，例如溶血素和白细胞素，以及小肽毒素，对红细胞和白细胞的裂解活性，尤其是代表针对金黄色葡萄球菌的主要防御层的中性粒细胞。这些毒素会导致组织损伤，促进远处器官的细菌传播和生长，并使病原体逃避宿主的先天免疫反应。此外，已知囊囊多糖（CP）可保护细菌免受多形核白细胞的吞噬活性，并已被验证为重要的疫苗靶标。综合生物治疗剂（IBT）正在追求与金黄色葡萄球菌感染的CP结合使用的基于毒素的疫苗。形成孔的？ - 蛋白酶（HLA），也称为？ - 毒素（AT），几乎由所有菌株产生，并且在几种金黄色葡萄球菌浸润性疾病中隐含。金黄色葡萄球菌细菌的人类最近的微生物学和血清学研究也表明了 - 毒素和苯酚可溶剂模量（PSM）对金黄色葡萄球菌致病性的重要性。根据我们最近的研究，针对HLA的抗体，toxin和psm？3显着降低了金黄色葡萄球菌菌的败血症的风险。该阶段I SBIR的目的是评估新型糖缀合物葡萄球菌毒素疫苗的可行性。设计了候选疫苗，它代表了HLA（AT62）N末端的关键结构结构域，该结构域（62）与–toxin和psm？3融合。我们先前已经表明，AT62疫苗可防止对细菌血症和肺炎的保护。该提案利用Glycovaxyn Inc.开发的一种独特而新颖的体内生物缀合技术，将这种杂交毒素与5型多糖类型（CP5）结合起来，作为生物轭效率的概念证明。生物缀合技术涉及用于表达弯曲杆菌PGLB的大肠杆菌细胞以及金黄色葡萄球菌CP8生物合成所需的基因。 PGLB将N连锁的CP8链转移到引入这些细胞中的疫苗构建体中。这项新技术是糖缀合物疫苗场的主要发展，其优势比常规化学共轭。在特定目的中，将产生三种毒素的1融合，并衰减识别并纳入融合构建体中。将测试候选疫苗的免疫原性，然后在AIM 2中产生的生物缀合物。成功完成了这项拟议的研究后，我们设想了II期SBIR，其中将为其他主要金黄色葡萄球菌囊囊多糖CP8和多价疫苗的功效（包括当前目前正在开发的我们组中正在开发的其他类似疫苗的疗效），在该研究中生产类似的生物缀合物）。最终目标是与大型制药公司建立战略合作伙伴关系，以将疫苗候选者转变为临床开发。