Proteomics-driven reverse vaccinology for gonorrhea.

 描述（由申请人提供）：淋病在全球范围内发病率很高，对全球生殖和新生儿健康产生重大影响。令人担忧的是，随着每一种用于淋病的新抗生素的引入，耐药性已经出现，包括对青霉素、四环素、氟喹诺酮类药物的耐药性，以及最近的第三代头孢菌素。治疗选择目前受到严重限制，淋病疫苗的开发是解决这一问题的关键，长期解决方案。然而，淋病疫苗的进展一直很缓慢，部分原因是淋病奈瑟菌（GC）中大量的表面分子经历了阶段或抗原变异，以及缺乏对保护性反应的了解。因此，淋病疫苗的开发可以受益于全面，公正的抗原发现方法。我们假设GC细胞包膜的许多成分是稳定的，对生物学功能很重要，因此代表了有吸引力的疫苗靶点。此外，有越来越多的证据表明，Th 1应答可以预防淋病，我们使用候选疫苗MtrE的初步数据支持Th 1诱导佐剂在诱导疫苗介导的保护中的有效性。因此，在这里，我们提出1）使用蛋白质组学指导的发现程序鉴定新的保守疫苗候选物。将进行高通量蛋白质组学研究，以扩大对不同感染部位遇到的刺激做出反应而表达的潜在抗原的阵列，包括：氧可用性（需氧和厌氧）、铁剥夺和人血清的存在; 2）评价作为疫苗候选物的选定蛋白质。将对19种候选蛋白质作为疫苗靶点的潜力进行严格评价，方法是使用当代GC临床分离物的不同集合来验证它们的表面暴露和保存，检查体外和实验性鼠感染期间的病理生理功能，并评估它们在小鼠中引发功能性抗体的能力; 3）确定候选抗原在鼠生殖道感染模型中的保护能力。将单独或彼此组合并与MtrE组合测试三种最有希望的抗原，以确定它们在鼠感染模型中针对GC的保护潜力。完成这些拟议的研究将为针对GC感染的预防性疫苗提供起点，并指导下一代疫苗设计，以及鉴定可能在GC发病机制中发挥作用的新型蛋白质。

 DESCRIPTION (provided by applicant): Gonorrhea occurs at high incidence worldwide and has a major impact on reproductive and neonatal health worldwide. Alarmingly, with each new antibiotic introduced for gonorrhea, resistance has emerged, including resistance to penicillin, tetracycline, fluoroquinolones, and recently the third-generation cephalosporins. Treatment options are currently seriously limited and the development of a gonorrhea vaccine is a critical, long- term solution to this problem. Progress on gonorrhea vaccines has been slow, however, in part due to the high number of surface molecules in Neisseria gonorrhoeae (GC) that undergo phase or antigenic variation and a lack of understanding of protective responses. Gonorrhea vaccine development can therefore benefit from a comprehensive, unbiased approach for antigen discovery. We hypothesize that many constituents of the GC cell envelope are stable and important for biological functions, and thus represent attractive vaccine targets. Moreover, there is growing evidence that Th1 responses protect against gonorrhea and we have preliminary data using the vaccine candidate MtrE that supports the effectiveness of Th1-inducing adjuvants in inducing vaccine-mediated protection. Accordingly, here we propose to 1) identify novel conserved vaccine candidates using a proteomics-guided discovery program. High-throughput proteomic studies will be performed to broaden the array of potential antigens that are expressed in response to stimuli encountered at different infection sites including: oxygen availability (aerobic and anaerobic), iron deprivation, and the presence of human serum; 2) evaluate the selected proteins as vaccine candidates. Nineteen protein candidates will be subjected to rigorous evaluation for their potential as vaccine targets by verifying their surface exposure and conservation using a diverse collection of contemporary GC clinical isolates, examining pathophysiological functions in vitro and during experimental murine infection, and by assessing their capacity to elicit functional antibodies in mice; 3) determine the protective capabilities of candidate antigens in a murine genital tract infection model. The three most promising antigens will be tested alone or in combination with each other and with MtrE, to determine their protective potential against GC in the murine infection model. Completion of these proposed studies will provide a starting point for preventive vaccine(s) against GC infections and guide next-generation vaccine design, as well as identify novel proteins that may play a role in GC pathogenesis.