A Rabbit Model of Epstein-Barr Virus Infection

EB病毒感染的兔模型

• 批准号: 10632071
• 负责人: Clare E Sample
• 金额: $ 20.15万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10632071
• 关键词: Adult; Animal Model; B lymphocyte immortalization; B-Lymphocytes; Biology; Burkitt Lymphoma; Cell Compartmentation; Cell Differentiation process; Cell Line; Cells; Cellular Structures; Complex; Development; EBV-associated disease; Epithelial Cells; Epithelium; Epstein-Barr Virus Infections; Epstein-Barr Virus Nuclear Antigens; Epstein-Barr Virus latency; Equilibrium; Genes; Genetic Transcription; Herpesviridae; Human; Human Herpesvirus 4; Immune; Immune response; Immune system; Immunocompetent; Immunologic Surveillance; Immunosuppression; In Vitro; Infection; Laboratories; Life Cycle Stages; Link; Mediating; Membrane Proteins; Memory B-Lymphocyte; Modeling; Nature; Oncogenic; Oryctolagus cuniculus; Phorbol Esters; Polymerase; Predisposition; Process; Productivity; Property; Proteins; Reaction; Recording of previous events; Reporting; Research; Research Personnel; Saliva; Sampling; Spleen; Structure of germinal center of lymph node; Surface Immunoglobulins; Therapeutic; Undifferentiated; Untranslated RNA; Vaccines; Viral; Viral Genome; Viral reservoir; Virus; Virus Diseases; Virus Replication; Work; adaptive immune response; antiviral drug development; cell mediated immune response; chronic infection; crosslink; design; experience; human pathogen; in vivo; laboratory rabbit; latency-associated protein; latent gene expression; latent infection; novel strategies; oral cavity epithelium; persistent EBV infection; prevent; programs; protein expression

PROJECT SUMMARY Epstein-Barr virus (EBV) is a herpesvirus that persists as a notable human pathogen with significant oncogenic potential that is related to its ability to establish lifelong latent infection within an immunocompetent host. Unlike for the vast majority of viruses, EBV-associated diseases are almost exclusively a consequence of latent infection instead of damage caused by virus replication. Latency poses significant problems, for example, in the development of effective anti-EBV therapeutics, since the majority of such compounds are designed to target viral polymerases and other proteins active only during virus replication; novel approaches are therefore needed to target latent EBV infection. Moreover, the ubiquitous nature of EBV (~95% infected by adulthood) and the highly evolved equilibrium that it has established within the human immune system, make development of an effective vaccine challenging. Both of these challenges could be greatly aided by a biologically accurate and tractable animal model of EBV infection, which currently does not exist. Surprisingly, several reports over the last decade have provided support for the laboratory rabbit as a model of EBV infection. In preliminary work, we have confirmed the most salient observations from these earlier studies, confirming that rabbits are indeed infectable, as indicated by persistence of the viral genome and expression of EBV genes associated with both the latent and productive stages of the virus lifecycle. However, several critical demonstrations are required to support authenticity of this model, and thus the extent to which it could be applied experimentally. In this application, we propose three specific aims to fill these gaps through complementary efforts by two senior investigators in the EBV field, and a third who has considerable expertise in the use of the rabbit as a model for virus infection and immune-related studies. Under Aim 1, Dr. Clare Sample will assess EBV infection in the epithelial cell component of the lifecycle, including the use of organotypic (raft) cultures of primary epithelium, a model she has developed and used to define EBV infection in human epithelium. Under Aim 2, Dr. Jeffery Sample will employ his expertise in the field of EBV latency to direct studies to elucidate whether establishment of a persistent infection in rabbit B cells is mechanistically comparable to the process that occurs in the human B-cell compartment. Finally, Dr. Neil Christensen, employing his expertise with the rabbit model will support work under Aims 1 and 2, and in Aim 3 will assess the humoral and cell-mediated immune responses to EBV infection in the rabbit, and whether they are comparable to key aspects of the adaptive immune response to EBV infection in the natural host. In summary, the proposed work will inform us whether the three basic host components that dictate EBV biology in its human host are equivalently functional in the rabbit: 1) Whether rabbit epithelium supports EBV replication that is critical for passage of the virus onto a naïve host; 2) Whether persistence of EBV in rabbit B cells is established through an authentic latent infection; and 3) Whether a comparable adaptive immune response is mounted to EBV infection in the rabbit.

项目概要 EB 病毒 (EBV) 是一种疱疹病毒，作为一种著名的人类病原体持续存在，具有显着的影响 致癌潜力与其在免疫功能健全的体内建立终生潜伏感染的能力有关 主持人。与绝大多数病毒不同，EBV 相关疾病几乎完全是以下原因造成的： 潜伏感染而不是病毒复制造成的损害。延迟会带来重大问题，例如， 开发有效的抗 EBV 疗法，因为大多数此类化合物旨在 靶向仅在病毒复制期间活跃的病毒聚合酶和其他蛋白质；因此，新颖的方法是 需要针对潜在的 EBV 感染。此外，EBV 的普遍性（约 95% 是在成年期感染的）和 它在人体免疫系统内建立的高度进化的平衡，使 有效的疫苗具有挑战性。这两个挑战都可以通过生物学上准确且可靠的方法得到极大的帮助。 EBV感染的易驯化动物模型，目前尚不存在。令人惊讶的是，多份报道称 过去十年为实验室兔子作为 EBV 感染模型提供了支持。在前期工作中，我们 已经证实了这些早期研究中最显着的观察结果，证实兔子确实是 病毒基因组的持久性和与两者相关的 EBV 基因的表达表明可感染 病毒生命周期的潜伏和生产阶段。然而，需要进行几次关键的演示 支持该模型的真实性，从而支持其实验应用的程度。在这个 在申请中，我们提出了三个具体目标，通过两位高级官员的互补努力来填补这些空白 EBV 领域的研究人员，以及第三位在使用兔子作为模型的方面拥有丰富专业知识的人 病毒感染和免疫相关研究。根据目标 1，Clare Sample 博士将评估 生命周期的上皮细胞组成部分，包括使用原代上皮的器官型（筏）培养物， 她开发并用于定义人类上皮中 EBV 感染的模型。在目标 2 下，Jeffery 博士 Sample 将利用他在 EBV 潜伏期领域的专业知识来指导研究，以阐明是否建立 兔子 B 细胞持续感染的过程在机制上与人类发生的过程相当 B 细胞室。最后，尼尔·克里斯滕森博士将利用他在兔子模型方面的专业知识来支持工作 目标 1 和 2 以及目标 3 将评估针对 EBV 感染的体液和细胞介导的免疫反应 以及它们是否与针对 EBV 感染的适应性免疫反应的关键方面具有可比性 在自然宿主中。总之，拟议的工作将告诉我们这三个基本主机组件是否 表明 EBV 在其人类宿主中的生物学特性在兔子中具有同等的功能： 1) 是否是兔子上皮细胞 支持 EBV 复制，这对于病毒传播至初始宿主至关重要； 2）是否持续存在 兔 B 细胞中的 EBV 是通过真正的潜伏感染建立的； 3) 是否具有可比的适应性 兔子对 EBV 感染产生免疫反应。