Structure and Function of Proxvirus Immune Evasion Domains

Proxvirus 免疫逃避结构域的结构和功能

• 批准号: 9012755
• 负责人: Daved H. Fremont
• 金额: $ 35.62万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9012755
• 关键词: Antibodies; Attenuated; Binding; Biological Assay; CXC Chemokines; Cell Surface Receptors; Cell surface; Cellular Assay; Chemotaxis; Clinical; Collaborations; Communicable Diseases; Communities; Complement; Complex; Cowpox; Cowpox virus; Crystallography; Data; Elements; Engineering; Ensure; Epitopes; Exhibits; Family; Fc Receptor; Flow Cytometry; GAG Gene; Goals; Host Defense; Immune; Immunologic Surveillance; In Vitro; Infection; Infection Control; Inflammation; Inflammatory Infiltrate; Integration Host Factors; Investigation; Kinetics; Knock-out; Ligand Binding; Ligands; Mass Spectrum Analysis; Mediating; Methods; Modeling; Mus; Orthopoxvirus; Pathogenesis; Phenotype; Play; Poxviridae; Process; Production; Property; Proteins; Publishing; Receptor Signaling; Recombinant Chemokine; Recombinants; Reporting; Resolution; Role; SCP 1; Series; Side; Site; Specificity; Structure; Surface Plasmon Resonance; T-Lymphocyte; Testing; Thermodynamics; Variant; Viral; Viral Pathogenesis; Virus; Virus Diseases; base; beta-Chemokines; chemokine; chemokine receptor; cytokine; expression vector; improved; in vitro Assay; mouse model; novel; pathogen; programs; protein structure; receptor; receptor binding; receptor mediated endocytosis; release of sequestered calcium ion into cytoplasm; research study; scaffold; screening; stoichiometry

Project 2: Structure and function of poxvirus immune evasion domains. In this project we will explore the functional and mechanistic attributes of a large family of sequence diverse proteins encoded by the zoonotic orthopoxvirus cowpox. We hypothesize that ten proteins In cowpox share a similar structural scaffolding that we have termed the poxvirus immune evasion (PIE) domain. Certain PIE proteins are known to sabotage host Immune surveillance and effector functions, and our goal Is to systematically determine the ligands and receptors for all ten PIE proteins encoded by the Brighton Red strain of cowpox. These efforts will be complemented by crystallographic investigations and quantitative binding assays to determine PIE protein structure and specificity, and structure-based engineering to develop variants with altered ligand/receptor Interactions. We will also undertake extensive In vitro functional studies of the PlEs, evaluating their mechanisms of action In the context of well-established Immunological assays. Our investigations will also be considered In the broader context of the battlefield of viral infection and host defense. In close collaboration with the Yokoyama and Fruh labs we will evaluate the pathogenesis of cowpox with either selective PIE proteins knocked out or their ligand/receptor binding determinants altered. The proposed experiments will result In a significantly improved functional annotation of cowpox and yield important clues about host-pathogen Interactions that are likely to benefit both the scientific and clinical communities.

项目2：痘病毒免疫逃逸结构域的结构和功能。在这个项目中，我们将探索由人畜共患正痘病毒牛痘编码的一个大家族的序列多样性蛋白的功能和机制属性。我们假设牛痘中的十种蛋白质具有相似的结构支架，我们将其称为痘病毒免疫逃避（PIE）结构域。已知某些PIE蛋白会破坏宿主的免疫监视和效应器功能，我们的目标是系统地确定所有十种PIE蛋白的配体和受体，这些蛋白由牛痘的布莱顿红病毒株编码.这些努力将通过晶体学研究和定量结合测定来补充，以确定PIE蛋白的结构和特异性，以及基于结构的工程设计来开发具有改变的配体/受体相互作用的变体。我们还将进行广泛的体外功能研究的PLEs，评估其作用机制的背景下，完善的免疫学测定。我们的调查也将被认为是在病毒感染和宿主防御的战场更广泛的背景下。在与横山和Fruh实验室的密切合作中，我们将评估牛痘的发病机制，无论是选择性PIE蛋白质敲除或其配体/受体结合决定簇改变。拟议的实验将显着改善牛痘的功能注释，并产生有关宿主-病原体相互作用的重要线索，这可能使科学界和临床界受益。