Antibody therapeutics for human viral hemorrhagic fevers and prevention of late neurological syndromes

项目总结 在东半球传播的丝状病毒和几种能够 在西半球的循环导致人类出血热，病例死亡 税率(15%至90%)。大多数人缺乏治疗选择。这里提出的研究项目是 一项利用病毒性出血热幸存者的血液来了解 在人类宿主和病原体方面的分子特征，使输注 抗体(被动免疫)在治疗人类病毒性出血中或多或少有效 发烧。为了实现这一目标，我们建议将人类抗体反应与 丝状病毒和天敌病毒。我们之所以选择比较这两组病毒，是因为 Junín病毒感染，输注幸存者血浆高效且常规使用 在临床上，它将感染的病死率降低到1%以下。相比之下， 虽然被动免疫在对抗埃博拉病毒的动物模型中也显示出了希望， 特别是在使用一种名为ZMapp的三抗体鸡尾酒的情况下，它还有待于 清楚地证明在人类身上是有效的。我们招募了朱尼恩病毒的幸存者， 埃博拉病毒和马尔堡病毒感染献血。在目标1中，我们将恢复 编码对虫媒病毒和丝状病毒表面起反应的抗体的基因 这些幸存者血液中的糖蛋白(抗体的靶标)。与 抗体基因序列，我们将产生大量的每一种已鉴定的抗体 作为重组蛋白，并在结合和病毒中和试验中进行研究。在AIM 2，我们将利用X射线结晶学和冷冻技术来确定抗体-糖蛋白的结构。 电子显微镜(低温电子显微镜)。在目标3中，我们将研究抗体的非中和性 在细胞检测和小动物模型中的抗病毒活性与我们的 合作者。我们还将确定我们是否可以减轻被动的已知副作用 对Junín病毒的免疫力-在接受治疗的幸存者中，10%的人晚期出现 可能是由于中枢神经系统中的病毒复制而导致的神经综合征 神经系统。我们研究的基本发现将帮助我们设计下一个 针对新出现的病毒产生被动免疫策略，这些病毒导致严重 人类疾病--也就是，特别定制的单抗疗法，其中 已知活性的纯化抗体也包括在内。

PROJECT SUMMARY Filoviruses that circulate in the Eastern Hemisphere and several arenaviruses that circulate in the Western hemisphere cause human hemorrhagic fevers with case fatality rates (15% to 90%). Most lack treatment options. The research project proposed here is an effort to use the blood of survivors of viral hemorrhagic fevers to understand the basic molecular features, on both the human host and pathogen side, that make transfusion of antibodies (passive immunity) more or less effective in treating human viral hemorrhagic fevers. To achieve this goal, we propose to compare the human antibody response to filoviruses and arenaviruses. We chose to compare both groups of viruses because for Junín virus infection, transfusion of survivor plasma is highly effective and routinely used clinically – it decreases the case fatality rate of infection to less than 1%. In contrast, while passive immunity has also shown promise in animal models a®gainst Ebola virus, particularly with the use of a three antibody-cocktail called ZMapp , it has yet to be clearly demonstrated as effective in humans. We have recruited survivors of Junín virus, Ebola virus, and Marburg virus infections for blood donation. In Aim 1, we will recover the genes encoding antibodies that react against the arenavirus and filovirus surface glycoprotein (the target of antibodies) from the blood of these survivors. With the sequences of antibody genes, we will produce large quantities of each identified antibody as recombinant proteins and study these in binding and viral neutralization assays. In Aim 2, we will determine antibody-glycoprotein structure using X-ray crystallography and cryo- electron (Cryo-EM) microscopy. In Aim 3, we will study the antibodies' non-neutralizing antiviral activity in cell-based assays and in small animal models working closely with our collaborators. We will also determine if we can alleviate a known side effect of passive immunity against Junín virus – development, in 10% of treated survivors, of a late neurological syndrome that probably occurs because of viral replication in their central nervous system. The basic findings from our research will help us design the next generation of passive immunity strategies against emerging viruses that cause severe human diseases – that is, specifically tailored monoclonal antibody therapies in which purified antibodies of known activity are included.