A Rabbit Model of Epstein-Barr Virus Infection

项目总结 爱泼斯坦-巴尔病毒(EBV)是一种疱疹病毒，作为一种重要的人类病原体持续存在，具有显著的 致癌潜力与其在免疫活性体内建立终身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由 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， 在开发有效的抗EBV疗法方面，因为大多数此类化合物旨在 靶向病毒聚合酶和其他仅在病毒复制期间活跃的蛋白质；因此，新的方法 需要针对潜在的EBV感染。此外，EBV的无处不在的性质(~95%由成年期感染)和 它在人类免疫系统内建立的高度进化的平衡，使 一种具有挑战性的有效疫苗。这两个挑战都可以很大程度上得到生物学上准确和 易驯服的EBV感染动物模型，目前尚不存在。令人惊讶的是，几份关于 过去十年为实验室兔作为EBV感染的模型提供了支持。在前期工作中，我们 证实了这些早期研究中最突出的观察结果，证实了兔子确实是 传染性的，从病毒基因组的持久性和与两者相关的EBV基因的表达来表明 病毒生命周期的潜伏期和产生期。然而，需要几个关键的演示来 支持这个模型的真实性，从而支持它可以在实验中应用的程度。在这 应用程序，我们提出了三个具体目标，通过两位资深员工的互补努力来填补这些空白 EBV领域的研究人员，以及在将兔子作为模型使用方面具有相当专业知识的第三人 病毒感染和免疫相关研究。根据目标1，Clare博士样本将评估EBV感染在 生命周期中的上皮细胞组成部分，包括使用原代上皮的器官型(RAFT)培养， 她开发的模型用于定义人类上皮中的EBV感染。在《目标2》中，杰弗里博士 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 在兔B细胞中持续感染的过程在机械上与在人类中发生的过程相似 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这项工作 在目标1和目标2中，以及在目标3中，将评估对EBV感染的体液和细胞介导的免疫反应 以及它们是否可与EBV感染的获得性免疫反应的关键方面相媲美 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 说明EBV在人类宿主中的生物学在兔身上具有同等的功能：1)兔上皮细胞 支持EBV复制，这对病毒传播到幼稚的主机上至关重要；2)是否持续 兔B细胞中的EBV是通过真实的潜伏感染建立的；以及3)是否有类似的适应性 兔对EB病毒感染产生免疫应答。

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.