Characterization of a novel rabbit herpes simplexvirus: a possible model for unde

描述(申请人提供)：阿拉斯加安克雷奇附近一个农场的55只兔子中，有一半以上感染了一种被称为LHV-4的疱疹病毒。大约28%的受影响动物因病情严重而死亡或被实施安乐死。感染兔的皮肤提取物在培养的兔皮肤和肾脏细胞以及Vero细胞中产生合胞体和细胞裂解。用简并引物保守的疱疹病毒基因进行聚合酶链式反应从纯化的病毒DNA中扩增序列。这些产物的测序允许设计特定的引物，从而可以确定一些基因的完整序列数据。对序列数据的分析表明，该病毒是单纯疱疹病毒属的成员，与牛疱疹病毒2、单纯疱疹病毒1、单纯疱疹病毒2和头猴疱疹病毒关系最近。新西兰大白兔实验性暴露于该病毒后，会导致严重的眼部疾病以及脾和淋巴结的坏死。此外，在急性感染期间(感染后5d内)，在多种组织中均可检测到病毒DNA，但只有在感染后14d才能在三叉神经节(TG)中持续检测到病毒DNA。瑞士韦伯斯特小鼠以1X103PFU/只眼暴露于该分离物，导致轻度眼部疾病和发烧。急性感染时在脾组织中检测到病毒DNA，感染20d后在TG中检测到病毒DNA。甲型疱疹病毒的一个独特特征是它们潜伏在TG中，除了潜伏相关的转录基因(LAT)外，没有其他基因表达。LAT在潜伏期中的作用仍不是很清楚。LAT基因可能是了解HSV-1或HSV-2潜伏再激活机制的关键。这项建议中概述的研究目的是研究这种新型疱疹病毒的LAT基因。为了完成这一点，我们将对LHV-4的基因组进行测序和注释，并确定LAT区域的特征。由于它与灵长类病毒谱系的密切关系，以及它感染兔子的能力，关于这种病毒潜伏期的信息可能为理解人类疱疹病毒感染的潜伏期提供一个自然的宿主模型系统。公共卫生相关性：在这项研究中，我们建议对最近发现的一种与HSV-1和HSV-2密切相关的兔单纯疱疹病毒病原体的基因组进行测序。本研究的目标是表征潜伏期相关转录基因(LAT)区域，并在自然宿主模型系统中研究LAT在潜伏期中的作用。。

DESCRIPTION (provided by applicant): A herpesvirus, referred to as LHV-4, was found to be associated with a disease affecting over half of the 55 rabbits on a farm near Anchorage, Alaska. About 28% of the affected animals died or were euthanized because of the severity of the illness. Extracts from the skin of infected rabbits produced syncytia and cell lysis in cultured rabbit skin and kidney cells, and in Vero cells. PCR using degenerative primers to conserved herpesvirus genes was used to amplify sequences from purified viral DNA. Sequencing of these products allowed the design of specific primers so that complete sequence data for a number of genes could be determined. Analysis of this sequence data indicates that the virus is a member of the simplex genus of herpesviruses and is most closely related to bovine herpesvirus 2, HSV-1, HSV-2, and cercopithecine herpesvirus. Experimental exposure of New Zealand White rabbits to the isolate resulted in severe eye disease and necrosis of the spleen and lymph nodes. In addition, viral DNA was identified in a variety of tissues by PCR during the acute infection (within 5 days post- infection), but was only detectable consistently in trigeminal ganglia (TG) after 14 days post- infection. Experimental exposure of Swiss Webster mice to the isolate at 1X103 PFU/eye resulted in mild eye disease and fever. Viral DNA was detected in spleen tissue during the acute infection, and was detectable in the TG after 20 day post-infection. One of the unique features of alphaherpes virus is that they become latent in the TG with no gene expression, except for the latency associated transcript gene (LAT). The role of LAT during latency is still not well understood. The LAT gene may be the key to understand the mechanism of HSV-1 or HSV-2 latency reactivation. The purpose of the research outlined in this proposal is to investigate LAT gene of this novel herpesvirus. To accomplish this we will sequence and annotate the genomes of LHV-4 and characterize the LAT region. Because of its close relationship to the primate virus lineage, and its ability to infect rabbits, information on latency in this virus may provide a natural host model system for understanding latency in herpesvirus infections of humans. PUBLIC HEALTH RELEVANCE: In this research application we propose to sequence the genome of a recently discovered Simplex herpesvirus pathogenic for rabbits that is closely related to HSV-1 and 2. The goal of this research is to characterize the latency associated transcript gene (LAT) region and investigate the role of LAT during latency in a natural host model system. .