A Rabbit Model of Epstein-Barr Virus Infection

EB病毒感染的兔模型

• 批准号: 10527462
• 负责人: Clare E Sample
• 金额: $ 24.21万
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10527462
• 关键词: Adult; Animal Model; Antiviral Agents; B lymphocyte immortalization; B-Lymphocytes; Biology; Burkitt Lymphoma; Cell Compartmentation; Cell Differentiation process; Cell Line; Cells; Cellular Structures; Complex; Development; EBV-associated disease; Epithelial; Epithelial Cells; 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; Link; Mediating; Membrane Proteins; Memory B-Lymphocyte; Modeling; Nature; Oncogenic; Oryctolagus cuniculus; Phorbol Esters; Polymerase; Process; 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.

项目摘要 爱泼斯坦 - 巴尔病毒（EBV）是一种疱疹病毒，一直是一种显着的人类病原体 与其在免疫能力内建立终身潜在感染的能力相关的致癌潜力 主持人。与绝大多数病毒不同，与EBV相关的疾病几乎完全是 潜在感染而不是病毒复制引起的损害。潜伏期有重大问题，例如 在开发有效的抗EBV疗法时，由于大多数此类化合物旨在 仅在病毒复制期间活跃于病毒聚合酶和其他蛋白质；因此，新颖的方法是 需要靶向潜在的EBV感染。此外，EBV的无处不在（〜95％被成年感染）和 它在人类免疫系统中建立的高度发展的平衡，使 有效的疫苗挑战。这两种挑战都可以通过生物学上的精确性来帮助 EBV感染的可处理动物模型，目前不存在。令人惊讶的是，关于 最近的十年为实验室兔子提供了支持，作为EBV感染的模型。在初步工作中，我们 已经证实了这些早期研究中最显着的观察结果，证实了兔子确实是 可感染，如病毒基因组的持续性和与两者相关的EBV基因的表达所表明 病毒生命周期的潜在和生产阶段。但是，需要几个关键示威 支持该模型的真实性，因此可以通过实验应用它的程度。在这个 申请，我们提出了三个特定的目的，以填补这些空白，通过两个高级的互补工作 EBV领域的调查人员和第三个在使用兔子作为模型的专业知识的第三名 病毒感染和免疫相关研究。在AIM 1下，Clare博士样本将评估EBV感染 生命周期的上皮细胞成分，包括使用原代上皮的有机（筏）培养物 她已经开发并用于定义人类上皮中的EBV感染。在AIM 2下，Jeffery博士 样本将利用他在EBV潜伏期领域的专业知识来指导研究以阐明是否建立 兔B细胞中持续感染的持续感染与人类发生的过程相媲美 B细胞室。最后，尼尔·克里斯滕森（Neil Christensen）博士在兔子模型中使用他的专业知识将支持工作 在目标1和2下，在AIM 3中将评估对EBV感染的体液和细胞介导的免疫调查 在兔子中，以及它们是否与对EBV感染的自适应免疫反应的关键方面相媲美 在天然主机中。总而言之，拟议的工作将告知我们是否三个基本主机组件是 在其人类宿主中指示EBV生物学在兔子中同样有效：1）兔上皮是否是否 支持EBV复制，这对于将病毒传递到幼稚的宿主至关重要。 2）是否持续 兔B细胞中的EBV是通过真实的潜在感染建立的。 3）是否具有可比的适应性 免疫反应安装在兔子中的EBV感染中。