Proteomics-Driven Reverse Vaccinology for Gonorrhea

摘要 淋球菌(Ng)，导致性传播感染的革兰氏阴性细菌 淋病被归类为研究和开发工作的高度优先病原体。在美国，超过50%的人 的Ng分离株对至少一种抗菌素具有耐药性，疾控中心已将Ng列为五大紧急情况之一 抗生素抗药性威胁。淋病的后果在性、性别、种族和民族方面尤其具有破坏性。 少数群体和资源有限的国家。吴氏超级细菌的地位，其高发病率，以及严重的健康和 淋病的心理影响使疫苗的开发成为必要。为了满足人们对 为了有效和安全地预防淋病，我们建议使用一种强大的病毒样颗粒(VLP)疫苗 具有高效分裂蛋白结合系统的平台，将有希望的Ng抗原(Ag)作为完整的- 长度和天然折叠的蛋白质。我们选择了六种有希望的脂蛋白作为这项建议的依据：1) 全球5K测序Ng分离株的特殊保护；2)表面暴露；3)诱导能力 杀菌抗体(Abs)；4)不同地理和时间差异的Ng株在不同时期的表达 淋病小鼠模型的体外条件和Ng感染；5)在Ng发病机制中的重要作用 和生理学。我们将：1)设计和生产抗原-VLP制剂；2)鉴定Ag-VLP和佐剂 产生强大免疫反应的组合；以及3)评估有希望的疫苗的效力和 阐明保护的免疫相关性。我们的方法在概念和技术方面进行了创新 淋病疫苗领域通过开创性的VLP展示保守的全长Ng脂蛋白，这将是 对有效性和潜在的保护机制进行比较评估。

ABSTRACT Neisseria gonorrhoeae (Ng), the gram-negative bacterium responsible for the sexually transmitted infection gonorrhea, is categorized as a high-priority pathogen for research and development efforts. In the US, over 50% of Ng isolates are resistant to at least one antimicrobial and the CDC has ranked Ng as one of the top 5 urgent antibiotic resistant threats. Gonorrhea outcomes are especially devastating in sexual, gender, racial, and ethnic minorities and resource-limited countries. Ng’s “superbug” status, its high morbidity, and the serious health and psychological impacts of gonorrhea necessitate vaccine development. To address the urgent need for an effective and safe vaccine against gonorrhea, we propose to use a powerful Virus-like-Particle (VLP) vaccine platform with a highly effective split-protein conjugation system to deliver promising Ng antigens (Ag) as full- length and natively folded proteins. We selected for this proposal six promising lipoproteins based on their: 1) exceptional conservation in >5K sequenced Ng isolates worldwide; 2) surface-exposure; 3) ability to elicit bactericidal antibodies (Abs); 4) expression in geographically and temporally diverse Ng strains, during different in vitro conditions and Ng infection in the gonorrhea mouse model; and 5) important functions in Ng pathogenesis and physiology. We will: 1) Design and produce antigen-VLP formulations; 2) Identify Ag-VLP and adjuvant combinations that generate robust immune responses; and 3) Evaluate efficacy of promising vaccines and elucidate the immune correlates of protection. Our approach presents conceptual and technical innovations in the gonorrhea vaccine field by pioneering VLP-display of conserved, full-length Ng lipoproteins, which will be comparatively assessed for efficacy and potential mechanisms of protection.