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

基本信息

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
项目类别：
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

项目摘要

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 Powassan病毒（POWV）是tick污染的黄病毒，会导致严重的脑炎，脑膜炎和长期神经损害。尽管POWV感染相对罕见，但该病毒分布广泛在常见载体（例如ixodes capapularis）（鹿tick虫）和报告中报告的病例的数量我们每年都在上升。没有批准的POWV感染疫苗或治疗方法。目标的目标 R21提案是两个方面。在AIM 1中，我们将研究蛋白质纳米颗粒免疫原的潜力重组表达的POWV糖蛋白E域III（EDIII）诱导中和抗体小鼠的反应。 EDIII是黄病毒亚基疫苗设计的有吸引力的目标，因为它是相对的小（约80个残基），并包含针对多种黄病毒（例如）的保护性抗体的表位登革热病毒（DENV），寨卡病毒（ZIKV），西尼罗河病毒（WNV）和循环病毒病毒（LIV）。但是，Ediii 由于单体不良免疫原性，因为它缺乏交联B细胞受体的交联能力（BCR）刺激稳健的抗体反应。可以通过在多价格式作为蛋白质纳米颗粒的一部分。我们已经产生了原型POWV EDIII 纳米颗粒疫苗使用间谍捕获器/间谍结合技术和试验研究表明该纳米颗粒可以引起小鼠中和抗体的中和抗体。我们将进一步优化这一点和相关的POWV 纳米颗粒疫苗。在AIM 2中，我们将隔离大型人类单克隆抗体（HU-MABS）来自单个B细胞分选的POWV感染的活着幸存者。 Hu-mab具有强大的潜力免疫疗法是因为它们高度具体，并且其人脚手架可最大程度地减少抗药物的风险抗体反应。此外，分析HU-MABS提供有关人免疫的直接信息然后可以用来告知疫苗设计的响应。我们将测试POWV HU-MABS的能力结合POWV E并中和POWV报告基因病毒颗粒（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。