Nanodisc-displayed Protein Vaccines

摘要 淋病奈瑟菌(Ng)是一种人类特有的病原体，是淋病的病原体，是一种性传播疾病。 传播感染给全球造成重大的健康负担，每年新增约7800万例。虽然经常 无症状、未经治疗的淋病可导致盆腔炎、宫外孕、不孕和 艾滋病毒传播/获得性增加。由于抗生素耐药性的不可阻挡的增加， 保护性淋病疫苗可能是未来控制疾病传播的唯一途径。最近的 B群脑膜炎奈瑟氏菌MenZB和4CMenB外膜囊泡疫苗的研制 为研制有效的淋病疫苗提供了有力的前提。回溯数据显示， 在免疫队列中，MenZB疫苗对淋病的有效率为31%。为了支持这一发现，我们 已经表明，用类似的4CMenB疫苗免疫显著增加了小鼠对Ng的清除 淋病模型和4CMenB疫苗免疫小鼠的血清与来自Ng的mtre、Bama和PilQ发生交叉反应 外膜。基于这些数据和表面曝光、无所不在、序列守恒和 在MTRE、BAMA和PilQ的重要细胞功能中的重要性，我们假设抗体针对 这些抗原的胞外区将提供对淋病的保护。因此，最重要的是 这个合作翻译项目的目标是开发一种针对MTRE、BAMA和MTRE的淋病疫苗(S) PilQ。亚单位抗原因其安全性、成本效益、 和快速准备。为了开发有效的淋病疫苗(S)，我们提出了一种创新的方法 将上述抗原加入到称为纳米盘(NDS)的纳米颗粒平台中并结合 在我们的疫苗配方中使用不同的佐剂。NDS将使抗原具有多价性和天然形状 是疫苗效力和效力的重要决定因素，而佐剂将用于增强强健 抗原特异性反应。淋病疫苗合作研究中心(GV CRC)项目3， 我们将：i)纯化MTRE、BAMA和PilQ的全长和/或β桶区域，并自然展示这些蛋白质 在NDS(特异性目标1)中；ii)蛋白质-NDS将与不同的佐剂成分结合以诱导健壮 和平衡的Th1/Th2反应，由此产生的血清将被评估免疫球蛋白亚型，血清 杀菌和吞噬细胞活性，并与完整的Ng结合(特定目标2)；以及iii)测试最多 抗原-ND/佐剂组合在Ng下、上生殖道小鼠模型中的应用前景 感染，以及在Ng/衣原体混合感染模型中，因为它们能够减少时间 感染的风险(具体目标3)。外膜将极大地促进GV CRC的成功 囊泡和蛋白质组学核心(核心B)、宿主反应监测核心(核心C)、功能性抗体 研究核心(核心D)和小鼠免疫/挑战核心(核心E)，这两个核心完全纳入AIMS 2和3，用于测试新城疫抗原疫苗的效力。

ABSTRACT Neisseria gonorrhoeae (Ng) is a human-specific pathogen and the etiological agent of gonorrhea, a sexually transmitted infection with a significant global health burden of ~78 million new cases annually. While often asymptomatic, untreated gonorrhea can lead to pelvic inflammatory disease, ectopic pregnancy, infertility, and increased transmission/acquisition of HIV. Because of the inexorable increase in antibiotic resistance, a protective gonorrhea vaccine may be the only way to control disease transmission in the future. The recent successes of the MenZB and 4CMenB outer membrane vesicle vaccines for Group B N. meningitidis (Nm) provides a strong premise for development of an effective gonorrhea vaccine. Retrospective data suggested that the MenZB vaccine was 31% effective against gonorrhea in the immunized cohort. In support of this finding, we have shown that immunization with the similar 4CMenB vaccine markedly increased Ng clearance in the mouse model of gonorrhea, and sera from 4CMenB-vaccinated mice cross-reacted with MtrE, BamA, and PilQ from Ng outer membranes. Based on these data and the surface exposure, omnipresence, sequence conservation, and importance in vital cellular functions of MtrE, BamA, and PilQ, we hypothesize that antibodies directed at the extracellular regions of these antigens will provide protection against gonorrhea. Accordingly, the overarching goal of this collaborative translational project is to develop a gonorrhea vaccine(s) by targeting MtrE, BamA, and PilQ. Subunit antigens are proven candidates for vaccine development due to their safety, cost-effectiveness, and rapid preparation. To develop effective gonorrhea vaccine(s), we propose an innovative approach of incorporating the aforementioned antigens into nanoparticle platforms called nanodiscs (NDs) and combining with different adjuvants in our vaccine formulations. NDs will enable antigen multivalency and native shape that are important determinants of vaccine potency and efficacy, while adjuvants will be used to amplify robust antigen-specific responses. For Project 3 of the Gonorrhea Vaccine Cooperative Research Center (GV CRC), we will: i) purify full-length and/or the β-barrel regions of MtrE, BamA, and PilQ and natively display the proteins in NDs (Specific Aim 1); ii) protein-NDs will be combined with different adjuvant compositions to induce robust and balanced Th1/Th2 responses, and the resulting sera will be assessed for immunoglobulin subtypes, serum bactericidal and opsonophagocytolytic activity, and binding to intact Ng (Specific Aim 2); and iii) test the most promising antigen-ND/adjuvant combinations in the lower and upper reproductive tract mouse models of Ng infection, as well as in a mixed Ng/Chlamydia muridarum infection model, for their capacity to decrease the time of infection (Specific Aim 3). The success of the GV CRC will be greatly enhanced by the Outer Membrane Vesicles and Proteomics Core (Core B), Host Response Monitoring Core (Core C), the Functional Antibody Study Core (Core D), and the Mouse Immunization/Challenge Core (Core E), which are fully integrated into Aims 2 and 3, for testing the efficacy of the antigen-ND vaccines.