Eliciting and isolating neutralizing antibodies against Powassan virus

引发并分离针对波瓦桑病毒的中和抗体

• 批准号: 10290425
• 负责人: Jonathan R. Lai
• 金额: $ 25.2万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10290425
• 关键词: Address; Adjuvant; Animal Model; Antibodies; Antibody Response; Antigens; Arboviruses; Area; B-Cell Antigen Receptor; B-Lymphocytes; Binding; Black-legged Tick; Blocking Antibodies; Blood specimen; Borrelia burgdorferi; C-terminal; Canada; Case Study; Cell Separation; Cell membrane; Cells; Cessation of life; Chikungunya virus; Clinical; Contracts; Deer Tick; Dengue Virus; Development; Diagnosis; Diagnostic; Dose; Encephalitis; Endosomes; Epitopes; FDA approved; Facial paralysis; Fatigue; Ferritin; Fever; Flavivirus; Genomics; Geometry; Glycoproteins; Goals; Head; Headache; Hour; Human; Immune response; Immunization; Immunotherapy; Incidence; Infection; Institutional Review Boards; Living Donors; Mediating; Membrane; Meningitis; Methodology; Monoclonal Antibodies; Mus; Nervous System Trauma; Nervous system structure; Neurologic; New York; Ontario; Paralysed; Patients; Pharmaceutical Preparations; Pilot Projects; Powassan virus; Prevention; Prevention approach; Proteins; Protocols documentation; Recombinants; Reporter; Reporting; Research; Resources; Respiratory Failure; Science; Seizures; Services; Subunit Vaccines; Surface; Survivors; Symptoms; Tail; Technology; Testing; Tick-Borne Diseases; Tick-Borne Encephalitis Virus; Ticks; Time; Transmembrane Domain; Vaccinated; Vaccine Design; Vaccines; Viral; Virion; Virus; Virus Diseases; West Nile virus; Work; Zika Virus; acute infection; antimicrobial; base; crosslink; design; disorder prevention; effective therapy; flavivirus glycoprotein E; human monoclonal antibodies; immunogenic; improved; insight; lumazine; monomer; nanoparticle; nervous system disorder; neutralizing antibody; pre-clinical; prevent; repository; risk minimization; scaffold; stem; support tools; symptom treatment; therapeutic candidate; three dimensional structure; tick-borne flavivirus; tick-borne virus; transmission process; vaccine candidate; vaccine development; vaccine evaluation; vector; vector control

SUMMARY Powassan virus (POWV) is a tick-disseminated flavivirus that causes severe encephalitis, meningitis, and long-term neurological damage. Although POWV infections are relatively rare, the virus is widely distributed among common vectors such as Ixodes scapularis (the deer tick) and the number of reported cases in the US is rising each year. There are no approved vaccines or treatments for POWV infection. The goals of this R21 proposal are two-fold. In Aim 1, we will investigate the potential of protein nanoparticle immunogens bearing recombinantly-expressed POWV glycoprotein E domain III (EDIII) to induce neutralizing antibody response in mice. EDIII is an attractive target for flavivirus subunit vaccine design because it is relatively small (~80 residues) and contains epitopes of protective antibodies against multiple flaviviruses such as Dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV), and Looping Ill virus (LIV). However, EDIII as a monomer is poorly immunogenic because it lacks the capacity to crosslink surface B-cell receptors (BCRs) to stimulate a robust antibody response. This limitation can be overcome by presenting EDIII in multivalent format as part of a protein nanoparticle. We have generated a prototypic POWV EDIII nanoparticle vaccine using Spycatcher/Spytag conjugation technology, and pilot studies have demonstrated this nanoparticle can elicit neutralizing antibodies in mice. We will further optimize this and related POWV nanoparticle vaccines. In Aim 2, we will isolate a large panel of human monoclonal antibodies (hu-mAbs) from a living survivor of POWV infection by single B cell sorting. Hu-mAbs have strong potential as immunotherapies because they are highly specific and their human scaffold minimizes the risk for anti-drug antibody responses. Furthermore, profiling hu-mAbs provides direct information about human immune response that can then be used to inform vaccine design. We will test the POWV hu-mAbs for their capacity to bind POWV E and neutralize POWV reporter virus particles (RVPs). This work will provide new insights and candidates for prevention and treatment of POWV infection.

概括 波瓦桑病毒 (POWV) 是一种蜱传播的黄病毒，可引起严重的脑炎、脑膜炎和 长期神经损伤。尽管 POWV 感染相对罕见，但该病毒分布广泛 肩胛硬蜱（鹿蜱）等常见媒介以及报告的病例数 美国每年都在上涨。目前还没有批准用于 POWV 感染的疫苗或治疗方法。本次活动的目标 R21 提案有两个方面。在目标 1 中，我们将研究蛋白质纳米颗粒免疫原的潜力 带有重组表达的 POWV 糖蛋白 E 结构域 III (EDIII)，可诱导中和抗体 小鼠的反应。 EDIII 是黄病毒亚单位疫苗设计的一个有吸引力的目标，因为它相对 小（约 80 个残基），包含针对多种黄病毒的保护性抗体表位，例如 登革热病毒 (DENV)、寨卡病毒 (ZIKV)、西尼罗河病毒 (WNV) 和循环 Ill 病毒 (LIV)。然而，EDIII 作为单体，其免疫原性较差，因为它缺乏交联表面 B 细胞受体的能力 （BCR）刺激强烈的抗体反应。通过在中呈现 EDIII 可以克服此限制 作为蛋白质纳米颗粒一部分的多价形式。我们已经生成了原型 POWV EDIII 使用 Spycatcher/Spytag 缀合技术的纳米颗粒疫苗，初步研究已证明 这种纳米颗粒可以在小鼠体内引发中和抗体。我们会进一步优化这个以及相关的POWV 纳米颗粒疫苗。在目标 2 中，我们将分离大量人单克隆抗体 (hu-mAb) 通过单 B 细胞分选从 POWV 感染的活幸存者中获得。 Hu-mAb 具有强大的潜力 免疫疗法，因为它们具有高度特异性，并且其人体支架最大限度地降低了抗药物风险 抗体反应。此外，对 hu-mAb 进行分析可提供有关人体免疫的直接信息 然后可用于为疫苗设计提供信息。我们将测试 POWV hu-mAb 的容量 结合 POWV E 并中和 POWV 报告病毒颗粒 (RVP)。这项工作将提供新的见解 以及预防和治疗 POWV 感染的候选人。