Nanodisc-displayed Protein Vaccines

纳米盘展示的蛋白质疫苗

• 批准号: 9900728
• 负责人: Aleksandra Elzbieta Sikora
• 金额: $ 25.24万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9900728
• 关键词: Adjuvant; Antibiotic Resistance; Antibiotics; Antibodies; Antigen Targeting; Antigenic Variation; Antigens; Bacterial Physiology; Binding; Biochemistry; Biological Assay; Cell physiology; Chlamydia muridarum; Clinic; Collaborations; Coupled; Data; Development; Dose; Ectopic Pregnancy; Effectiveness; Epitopes; Etiology; Female; Fimbriae Proteins; Future; Goals; Gonorrhea; Growth; HIV; Human; Immune response; Immune system; Immunization; Immunize; Immunoglobulins; Immunology; Individual; Infection; Infertility; Lead; Length; Membrane; Meningitis; Metabolic Clearance Rate; Microbiology; Modeling; Monitor; Mus; Neisseria gonorrhoeae; Neisseria meningitidis; Pathogenesis; Pelvic Inflammatory Disease; Phase; Preparation; Production; Protein Inhibition; Proteins; Proteomics; Research; Resistance; Role; Safety; Scaffolding Protein; Serum; Sexually Transmitted Diseases; Shapes; Surface; Testing; Time; Vaccinated; Vaccine Research; Vaccines; Vagina; Vesicle; bactericide; base; capsule; cell killing; cohort; cost effectiveness; disease transmission; disorder control; efficacy testing; experimental study; extracellular; global health; gonorrhea vaccine; his6 tag; innovation; inter-institutional; lipid nanoparticle; mouse model; nanodisk; nanoparticle; novel; pathogen; reproductive tract; response; success; transmission process; trend; vaccine candidate; vaccine development

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.

摘要 淋病奈瑟菌（Neisseria gonorrhoeae，Ng）是一种人类特异性病原体，是淋病的病原体，是一种性传播疾病。 传播感染，每年约有7800万新病例，给全球造成重大健康负担。虽然经常 无症状，未经治疗的淋病可导致盆腔炎，宫外孕，不孕症， 增加艾滋病毒的传播/感染。由于抗生素耐药性的无情增加， 保护性淋病疫苗可能是未来控制疾病传播的唯一途径。近期 MenZ B和4 CM MenZ B外膜囊泡疫苗用于B N群的成功。脑膜炎（Nm） 为开发有效的淋病疫苗提供了有力的前提。回顾性数据表明， MenZB疫苗在免疫组群中对淋病的有效性为31%。为了支持这一发现，我们 已经显示用类似的4CMenB疫苗免疫显著增加小鼠中的Ng清除 淋病模型，以及来自4CMenB免疫小鼠的血清与来自Ng的MtrE、BamA和PilQ交叉反应 外膜根据这些数据和地表暴露，无所不在，序列保护， MtrE、BamA和PilQ在重要细胞功能中的重要性，我们假设针对MtrE、BamA和PilQ的抗体在细胞中的重要性。 这些抗原的细胞外区域将提供抗淋病的保护。因此，总体而言， 这个合作翻译项目的目标是通过靶向MtrE，BamA和 PilQ亚单位抗原由于其安全性、成本效益， 快速制备。为了开发有效的淋病疫苗，我们提出了一种创新的方法， 将上述抗原掺入称为纳米盘（ND）的纳米颗粒平台中， 在我们的疫苗配方中加入不同的佐剂。ND将使抗原多价性和天然形状， 是疫苗效力和效力的重要决定因素，而佐剂将用于增强疫苗的稳定性。 抗原特异性反应。对于淋病疫苗合作研究中心（GV CRC）的项目3， 我们将：i）纯化MtrE、BamA和PilQ的全长和/或β桶区，并天然展示这些蛋白质 ii）蛋白质-ND将与不同的佐剂组合物组合以诱导稳健的免疫应答; 和平衡的Th 1/Th 2应答，并且将评估所得血清的免疫球蛋白亚型、血清 杀菌和调理吞噬细胞活性，并结合完整的Ng（特异性目标2）;和iii）测试最多的 在Ng的下和上生殖道小鼠模型中有希望的抗原-ND/佐剂组合 感染，以及在混合的Ng/小鼠衣原体感染模型中，因为它们能够减少 感染（具体目标3）。GV CRC的成功将通过外膜大大增强 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/攻毒核心（核心E），完全整合到目标中 2和3，用于测试抗原-ND疫苗的效力。