Proteomics-driven reverse vaccinology for gonorrhea.

 描述（由申请人提供）：淋病在全世界范围内发病率很高，对全世界的生殖和新生儿健康产生重大影响。令人担忧的是，随着每种新的淋病抗生素的出现，耐药性也随之出现，包括对青霉素、四环素、氟喹诺酮类药物以及最近的第三代头孢菌素的耐药性。目前的治疗选择严重有限，而开发淋病疫苗是解决这一问题的关键、长期的解决方案。然而，淋病疫苗的进展缓慢，部分原因是淋病奈瑟菌 (GC) 中存在大量表面分子，这些分子会发生相变或抗原变异，并且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并且对生物功能很重要，因此代表了有吸引力的疫苗靶点。此外，越来越多的证据表明 Th1 反应可预防淋病，并且我们使用候选疫苗 MtrE 获得了初步数据，支持 Th1 诱导佐剂在诱导疫苗介导的保护方面的有效性。因此，我们在这里建议 1) 使用蛋白质组学指导的发现计划来鉴定新型保守候选疫苗。将进行高通量蛋白质组学研究，以扩大针对不同感染部位遇到的刺激而表达的潜在抗原的范围，包括：氧气可用性（需氧和厌氧）、缺铁和人血清的存在； 2) 评估所选蛋白质作为候选疫苗。 19种候选蛋白质将接受严格的评估，以确定其作为疫苗靶标的潜力，方法是使用多种当代GC临床分离株验证其表面暴露和保存情况，检查体外和实验性小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)确定候选抗原在小鼠生殖道感染模型中的保护能力。三种最有希望的抗原将单独进行测试，或者相互结合以及与 MtrE 联合测试，以确定它们在小鼠感染模型中针对 GC 的保护潜力。这些拟议研究的完成将为胃癌感染的预防性疫苗提供起点，指导下一代疫苗设计，并鉴定可能在胃癌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.