Proteomics-Driven Reverse Vaccinology for Gonorrhea

摘要 淋病奈瑟菌（Ng），革兰氏阴性细菌，负责性传播感染 淋病被归类为研究和开发工作的高优先级病原体。在美国，超过50% 的Ng分离株对至少一种抗菌药物具有耐药性，CDC已将Ng列为前5名紧急病例之一 抗生素耐药性威胁淋病的结果在性、性别、种族和民族方面尤其具有破坏性 少数民族和资源有限的国家。Ng的“超级细菌”地位，其高发病率，以及严重的健康和 淋病的心理影响需要疫苗的开发。为了解决迫切需要 有效和安全的淋病疫苗，我们建议使用强大的病毒样颗粒（VLP）疫苗 具有高效分裂蛋白缀合系统的平台，以将有前景的Ng抗原（Ag）作为完整的 长度和天然折叠的蛋白质。我们选择了六种有前途的脂蛋白，基于它们的：1） 在全世界> 5 K测序的Ng分离株中具有特殊的保守性; 2）表面暴露; 3）引发 杀菌抗体（Abs）; 4）在地理上和时间上不同的Ng菌株中的表达，在不同的 体外条件和淋病小鼠模型中的Ng感染;以及5）Ng发病机制中的重要功能 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 产生强有力的免疫应答的组合;以及3）评估有希望的疫苗的功效， 阐明保护的免疫相关性。我们的方法提出了概念和技术创新， 淋病疫苗领域的开拓性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了比较评估。

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.