Human antibody modification of enterovirus D68 infection

项目摘要/摘要 急性弛缓性脊髓炎(AFM)是一种类似脊髓灰质炎的儿童疾病，于2014年在美国出现。 从那时起，AFM每隔一年8月至10月爆发一次，与肠道病毒爆发同时发生 D68(EV-D68)感染。现在越来越多的证据表明，EV-D68是AFM的主要原因。EV-D68， 与脊髓灰质炎病毒属于同一肠道病毒属，主要引起呼吸道感染。然而， EV-D68的神经致病机制及人体免疫应答特点 人们对此了解甚少。这个为期五年的研究职业发展奖将提供培训和 培养PI建立独立研究计划所需的技能，重点是 了解EV-D68如何导致呼吸道和神经系统疾病以及人类抗体如何改变这一点 发病机制。现为儿科传染病终身制助理教授 以及北卡罗来纳大学教堂山医学院的微生物学和免疫学。他的 到目前为止，培训的重点是分离和鉴定人的单抗。 有EV-D68感染史的受试者。他将通过进一步的培训来补充这一经验，以开发 利用EV-D68感染的体内、小鼠和体外人组织模型研究其发病机制 病毒和人类抗体，包括单克隆和多克隆，如何改变发病机制。短期目标是 拟议的研究是为了检验人类抗体阻止EV-D68引起EV-D68的中心假设 AFM，但不能预防呼吸道疾病。国际和平倡议将由国家专家指导，研究 微小核糖核酸病毒(克雷格·卡梅隆博士和斯坦利·莱蒙博士)，人类呼吸道上皮细胞培养(雷蒙德博士 泡菜)、病毒感染的小鼠模型(马克·海斯博士)和人类抗体(小詹姆斯·克罗博士)。这个 具体目的是1)确定EV-D68特异性人类抗体如何改变病毒诱导的呼吸 疾病和AFM在小鼠感染模型中的作用，以检验抗体有效地阻止EV- D68通过防止传播到中枢神经系统而导致原子力显微镜；以及2)调查决定 抗体对体外分化的人呼吸道上皮细胞EV-D68感染的抑制作用 气-液界面，以检验抗体对初始呼吸道粘膜的保护作用不佳的假设 感染。总体而言，这些研究将有助于确定EV-D68中和抗体是否是一种机制 通过精确定位能够限制EV-1的抗体的数量和质量来保护AFM的相关性- D68呼吸道感染。行业合作者正在开发由PI AS制造的保护性单抗 人类治疗剂，这将是EV-D68儿童唯一可用的靶向治疗 感染。因此，这些研究直接转化为人类健康的重要性，除了有助于 对疾病有更基本的了解。

Project Summary/Abstract Acute flaccid myelitis (AFM) is a poliomyelitis-like illness of children that emerged in the US in 2014. Since then, AFM outbreaks occur August-October every other year, concurrent with outbreaks of enterovirus D68 (EV-D68) infection. Increasing evidence now indicates that EV-D68 is a principal cause of AFM. EV-D68, in the same Enterovirus genus as poliovirus, primarily causes respiratory tract infections. However, the mechanisms of EV-D68 neuropathogenesis and the characteristics of the human immune response to EV-D68 are poorly understood. This five-year research career development award will provide training and development of the skills necessary for the PI to establish an independent research program focused on understanding how EV-D68 causes respiratory and neurologic disease and how human antibodies modify this pathogenesis. Currently the PI is an Assistant Professor on the tenure track in Pediatric Infectious Diseases and Microbiology & Immunology at the University of North Carolina at Chapel Hill School of Medicine. His training to date has focused on the isolation and characterization of human monoclonal antibodies (mAbs) from subjects with prior EV-D68 infection. He will supplement this experience with further training in developing in vivo mouse and ex vivo human tissue models of EV-D68 infection with which to study the pathogenesis of this virus and how human antibodies, both monoclonal and polyclonal, modify pathogenesis. The short-term goal of the proposed studies is to test the central hypothesis that human antibodies prevent EV-D68 from causing AFM but do not protect against respiratory disease. The PI will be mentored by national experts in the study of picornaviruses (Drs. Craig Cameron and Stanley Lemon), human airway epithelial cultures (Dr. Raymond Pickles), mouse models of virus infection (Dr. Mark Heise), and human antibodies (Dr. James Crowe, Jr.). The specific aims are to 1) determine how EV-D68-specific human antibodies modify virus-induced respiratory disease and AFM in a mouse model of infection, to test the hypothesis that antibodies efficiently preclude EV- D68 from causing AFM by preventing dissemination to the CNS; and 2) investigate factors that determine the efficacy of antibody at inhibiting EV-D68 infection in differentiated primary human airway epithelial cells at an air-liquid interface, to test the hypothesis that antibodies poorly protect against initial respiratory mucosal infection. Overall, these studies will help determine whether EV-D68 neutralizing antibodies are a mechanistic correlate of protection from AFM by pinpointing the quantity and quality of antibodies capable of restricting EV- D68 infection to the respiratory tract. Industry collaborators are developing protective mAbs made by the PI as human therapeutic agents, which would be the only available targeted therapy for children with EV-D68 infection. Thus, these studies directly translate to importance in human health in addition to contributing to a more fundamental understanding of disease.