Structural and functional basis of ultra potent CHKV neutralization by human mAbs

人单克隆抗体超强中和 CHKV 的结构和功能基础

• 批准号: 8894218
• 负责人: James E Crowe
• 金额: $ 76.18万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-10 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8894218
• 关键词: Acute; Aedes; Affinity; Alphavirus; Americas; Antibodies; Antibody Response; Antigens; Antiviral Agents; Area; B-Lymphocytes; Binding; Biological Assay; Bos taurus structural-GP protein; Caribbean region; Cell Culture Techniques; Cell membrane; Characteristics; Chikungunya virus; Chronic Disease; Complement; Complement 1q; Complex; Country; Coupled; Coupling; Cryoelectron Microscopy; Culicidae; Disease; Disease Outbreaks; Donor Selection; Elements; Encephalopathies; Engineering; Epidemic; Epitopes; Fab Immunoglobulins; Fever; Foundations; Future; Generations; Genes; Genotype; Glycoproteins; Goals; Human; Immunocompromised Host; Immunoglobulin Somatic Hypermutation; Immunologics; Immunotherapeutic agent; Infection; Inflammatory; Interferometry; Knowledge; Lead; Location; Memory B-Lymphocyte; Molecular; Molecular Genetics; Monoclonal Antibodies; Mus; Mutagenesis; Mutate; Mutation; North America; Pattern; Polyarthralgias; Reaction; Relative (related person); Reporting; Research Personnel; Role; Siblings; Site; South America; Specificity; Staging; Structural Biologist; Structure; Structure of germinal center of lymph node; Surface; Surface Plasmon Resonance; Technology; Testing; Therapeutic Agents; Vaccines; Variant; Viral Antigens; Viral Hemorrhagic Fevers; Virion; Virus; Virus Diseases; Virus Replication; West Indies; X-Ray Crystallography; antigen binding; arthropathies; base; crosslink; design; env Gene Products; experience; human disease; human monoclonal antibodies; in vivo; innovation; mouse model; neutralizing monoclonal antibodies; next generation sequencing; novel vaccines; prevent; public health relevance; research study; screening; transmission process; vaccine development; vector; vector mosquito; virus envelope

 DESCRIPTION (provided by applicant): Chikungunya virus (CHIKV) is a re-emerging mosquito-borne alphavirus that cycles directly between humans and mosquitoes and causes a debilitating febrile illness in humans on a global scale. Autochthonous transmission in the New World now has been described, with outbreaks in at least 19 countries in the Americas and over a quarter million cases in just 6 months. CHIKV disease cases have been reported to ArboNET in travelers returning to the US from the Caribbean in 29 states, as of July 1. The potential for epidemics in North and South America is high due to the ubiquitous distribution of one of its primary vectors, Aedes albopictus. Despite the possibility for infecting and causing disease in millions, specific treatments or vaccines for CHIKV are not available. A primary goal of this project is to define the molecular, genetic, immunologic, and structural characteristics of ultra-potent neutralizing human mAbs with broad activity against all genotypes of CHIKV. Additional goals include defining the mechanistic correlates of protection by these ultra-potent neutralizing mAbs. In these studies, we will elucidate how antiviral Abs with exceptional inhibitory activity exert their action in cell culture and in vivo. The approach will include high efficiency isolationof human mAbs, coupled with innovative antibody gene repertoire studies based on nextgen sequencing. Several hypotheses will be tested, including the concept that ultra-potent neutralizing activity results from features of both the antibodies (extensive mutations due to persistent CHIKV infection) and the antigen (binding to quaternary epitopes on multiple adjacent envelope proteins and blocking structural transitions critical for virus entry or release). Althoug our focus is to understand how and why ultra-potent human mAbs inhibit CHIKV, the studies likely will be relevant to general principles of antibody neutralization of many different viruses. Beyond defining the molecular and structural basis of Ab neutralization of CHIKV, these studies will generate a group of fully human mAbs that can prevent and treat CHIKV infection and persistence in mice, which could be developed in the future as a possible combination immunotherapeutic for humans. Studies in this project, while targeted against CHIKV, likely will inform future Ab-based and/or vaccine efforts against other alphaviruses that cause human disease. We have assembled a unique group of investigators, including a human Ab expert, a molecular virologist with experience in Ab-virus interactions, and two accomplished structural biologists with specific expertise in alphaviruses, including CHIKV, to pursue these studies.

 描述（由适用提供）：基孔肯雅病毒（Chikv）是一种重新出现的蚊子 - 传播的α病毒，直接在人类和蚊子之间循环，并在全球范围内引起人类的衰弱的高温疾病。现在已经描述了新世界的自动传播，在美洲至少19个国家发生了爆发，在短短6个月内发生了超过25万个案件。据报道，据报道，截至7月1日，在29个州从加勒比海返回美国的旅行者中，Chikv疾病案件的可能性很高。由于其主要载体之一的无处不在，其主要媒介是其主要载体，而北美地区的流行潜力很高。尽管有可能感染并引起数百万次疾病，但CHIKV的特定治疗或疫苗尚无。该项目的主要目的是定义具有广泛活性对CHIKV所有基因型的广泛活性的超级中和人mAb的分子，遗传，免疫和结构特征。其他目标包括定义这些超动力中和的mAb的机械保护相关性。在这些研究中，我们将阐明具有特殊抑制活性的抗病毒ABS如何在细胞培养和体内发挥作用。该方法将包括高效率分离人mAb，并基于NextGen测序，再加上创新的抗体基因曲目研究。将测试几种假设，包括概念，即超动力中和活性由抗体的特征（由于持续性CHIKV感染而引起的广泛突变）和抗原（与多个邻接的内膜蛋白质和封闭的季节结合至关重要的季度表位，对病毒蛋白和释放至关重要）。 mAb抑制CHIKV，这些研究可能与许多不同病毒的抗体中和的一般原理有关。 除了定义CHIKV AB协商的分子和结构基础外，这些研究还将产生一组完全人类的单元单位，可以预防和治疗小鼠的Chikv感染和持久性，将来可以作为可能的人类的免疫治疗方法来开发。该项目的研究虽然针对CHIKV，但可能会为未来的基于AB的和/或疫苗的努力告知其他引起人类疾病的α病毒。我们组建了一组独特的研究者，其中包括人类AB专家，具有AB-VIRUS相互作用经验的分子病毒学家，以及两名成熟的结构生物学家，在包括CHIKV在内的α病毒中具有特定专业知识，以从事这些研究。