Project 2 - Vanderbilt University

项目摘要 亨尼帕病毒属副粘病毒科的RNA病毒属，包含五个已建立的种。 埃尼帕病毒在澳大利亚和亚洲的蝙蝠身上自然存在，最近在非洲也发现了这种病毒， 它们的寄主范围很广。这些人畜共患病的病原体在国内可导致严重的疾病和死亡。 动物和人类。人类亨德拉和尼帕病毒感染的预防和治疗选择 是有限的。抗Hendra/Nipah人单抗有望成为有价值的抗病毒治疗药物 人类亨德拉和尼帕病毒病的对策。在这里，我们将隔离天然的 出现与Hendra和Nipah病毒F或Npah病毒有交叉反应的人类单抗 G蛋白并中和这两种病毒。在初步实验中，我们已经分离出了一些表现出 在中和分析中具有很高的效力，这表明它们具有很高的预防和治疗潜力 人类的分子。我们在这里提出了一系列目标，这些目标将对发展做出重大贡献 与Hendra和Nipah病毒F和G糖蛋白反应的人源单抗的鉴定 为临床研究做准备。这项工作将确定并充分描述一组极有希望的 抗体，其目标是识别和选择先导化合物并推进其临床前研究 发展。这项工作有两个主要的具体目标：目标1)从患者中分离出人mAbs 以前感染过埃尼帕病毒或接触过埃尼帕病毒疫苗抗原。在这一目标中，人类单抗将 被鉴定为识别在霍尼帕病毒中保守的表位，并在 低浓度。将对接触亨德拉病毒马疫苗的受试者的血细胞进行病毒筛查 特定的抗体。这些细胞将被转化为稳定的人类杂交瘤细胞系，并受到高度的 筛选与Hendra和Nipah G蛋白结合的功能性抑制病毒的抗体 复制。目的2：研制治疗黑热病病毒感染的单抗。目标1中确认的抗体将是 测试了它们对所有埃尼帕病毒保守表位的广泛识别以及它们的能力 在培养中中和。然后将在多个动物模型中测试优先抗体的治疗效果 包括非人类灵长类在内的感染。将选择引线，并由Mapp制作CHO细胞系 用于抗体生产的生物制药，为cGMP生产和IND规划做准备。这项工作 承诺生产一种同类最好的抗体制剂，以广泛而有效地对抗埃尼帕病毒， 可用于治疗或预防人类埃尼帕病毒感染。

Project Summary The Henipavirus genus of RNA viruses in the family Paramyxoviridae contains five established species. Henipaviruses are found naturally in bats in Australia and Asia and more recently have been found in Africa, and they have a wide host range. These zoonotic pathogens can cause severe illness and death in domestic animals and humans. The prophylactic and therapeutic options for Hendra and Nipah virus infections in man are limited. Anti-Hendra/Nipah human mAbs are expected to be valuable antiviral therapeutics as countermeasures to Hendra and Nipah virus disease in humans. Here, we will isolate panels of naturally occurring human monoclonal antibodies (mAbs) that bind cross-reactively to both Hendra and Nipah virus F or G proteins and neutralize both viruses. In preliminary experiments, we have isolated some mAbs that exhibit very high potency in neutralization assays, suggesting they have high potential as prophylactic and therapeutic molecules for humans. We propose here a series of aims that will contribute significantly to the development and characterization of such human mAbs reactive to the F and G glycoproteins of Hendra and Nipah virus in preparation for clinical studies. The work will identify and fully characterize a panel of highly promising antibodies with the goal of identifying and selecting lead compounds and advancing their preclinical development. The work is organized in two major Specific Aims: Aim 1) Isolation of human mAbs from patients previously infected with henipavirus or exposed to henipavirus vaccine antigens. In this Aim, human mAbs will be identified that recognize epitopes that are conserved across henipaviruses and neutralize those viruses at low concentration. Blood cells from a subject exposed to Hendra virus equine vaccine will be screened for virus specific antibodies. These cells will be converted to stable human hybridoma cell lines and subjected to high- throughput screening to identify Abs that bind to Hendra and Nipah G proteins and functionally inhibit virus replication. Aim 2: Develop Abs for the treatment of henipavirus infections. Antibodies identified in Aim 1 will be tested for their broad recognition of conserved epitopes across all henipaviruses and for their ability to neutralize in culture. Prioritized antibodies then will be tested for therapeutic efficacy in multiple animal models of infection including nonhuman primates. The leads will be selected, and CHO cell lines will be made by Mapp Biopharmaceutical for Ab production, in preparation for cGMP manufacture and IND planning. The work promises to yield a best-in-class antibody preparation for broad and potent activity against henipaviruses that can be used to treat or prevent human henipavirus infections.