Theiler?s virus as a potential cause of Vilyuisk encephalitis

描述（由申请方提供）：本R21探索性提案的目的是确定V-1分离株（称为Vilyuisk人脑炎病毒（VHEV），可能代表泰勒病毒的一个单独分支）是否为Vilyuisk脑脊髓炎（VE）的病原体。虽然病毒性VE的原因仍然未知，但有令人信服的证据将VHEV与VE联系起来。VE已经从Vilyui河中游沿着的地方性地区蔓延到400公里，到达Lena-Aldan河谷的一个新的人口更多的地理区域，西伯利亚（俄罗斯）Sahka共和国首都雅库茨克位于该地区。由于航空旅行自由度的增加而导致的全球化可能导致VE进一步传播到世界其他地区。最近在来自北美和其他四大洲的不同地理位置的患有呼吸道和呼吸道疾病的儿童中鉴定出被称为藏红花病毒（SafV）的泰勒病毒，表明泰勒病毒是全球性分布的，并且通常感染儿童和可能的成人。泰勒病毒的出现是我们对VE病因重新产生兴趣的催化剂，因为泰勒病毒引起人类中枢神经系统（CNS）疾病是可信的。事实上，PI刚刚从欧洲儿童脑炎病例中了解到SafV分离。因此，推测泰洛病毒代表一组引起中枢神经系统CNS感染的新兴人类病毒是有趣的，包括VE（假设）。本研究的主要目的是：（1）为探讨VHEV与VE临床疾病之间的关系，我们将测定VE患者和对照组血清中VHEV的ELISA和中和抗体的滴度、血清与脑脊液的比值、VE脑脊液寡克隆区带（OCB）对VHEV的反应性，以及从VE脑脊液中提取的总RNA中是否能扩增出包含高度可变的VP 2 puff（EF）结构的320 bp的HBV扩增子。(2)在VHEV不能与VE连接的情况下，从VE CSF提取的总RNA的逆转录cDNA将与寡核苷酸病毒微阵列（GreenChipsPm）杂交，并用于高通量测序，以通过减去人转录体来检测特定脊椎动物病毒家族的外源（非人）序列。公共卫生相关性：这是一个探索性的建议，以确定发生在俄罗斯东北部（西伯利亚）的萨哈卡共和国的慢性中枢神经系统疾病的病毒原因。这种疾病，Vilyuisk脑脊髓炎，与多发性硬化症有一些相似之处，似乎是由人类泰洛病毒引起的。鼠对应物Theiler鼠脑脊髓炎病毒为多发性硬化症提供了高度相关的实验动物模型。

DESCRIPTION (provided by applicant): The goal of this R21 exploratory proposal is to determine whether the V-1 isolate, termed Vilyuisk human encephalitis virus (VHEV), which may represent a separate clade of theilovirus, is the causative agent of Vilyuisk encephalomyelitis (VE). Although the viral cause of VE remains unknown compelling evidence links VHEV to VE. VE has spread 400 km from the area of endemicity along the middle reaches of the Vilyui River to a new more populous geographic area in the Lena-Aldan river valleys where Yakutsk, the capital of the Sahka Republic in Siberia (Russia), is located. Globalization due to increased freedom of air travel could result in further spread of VE to other regions of the world. Recent identification of theiloviruses designated Saffold viruses (SafV) in children with respiratory and diarrheal diseases from separate geographic locations in North America and four other continents indicate that theiloviruses are global in distribution and commonly infect children and possibly adults. The emergence of theiloviruses is the catalyst for our renewed interest in the cause of VE since it is creditable that theiloviruses cause human central nervous system (CNS) diseases. In fact, the PI has just learned of SafV isolations from cases of encephalitis in children in Europe. Therefore, it is intriguing to speculate theiloviruses represent an emerging group of human viruses that cause central nervous system CNS infections, including VE (hypothesis). Two specific aims are proposed: (1) To investigate a link between VHEV and the clinical disease of VE, we will determine serum titers and serum:CSF ratios of ELISA and neutralizing antibodies to VHEV in VE cases and controls, VE cerebrospinal fluid (CSF) oligoclonal band (OCB) reactivity to VHEV, and whether a 320- bp Cardiovirus amplicon that includes the highly variable VP2 puff (EF) structures can be amplified from total RNA extracted from VE CSF. (2) In the event that VHEV cannot be linked to VE, reverse transcribed cDNA from total RNA extracted from VE CSF will be hybridized to oligonucleotide viral microarrays (GreenChipsPm) and also used for high-throughput sequencing to detect foreign (non-human) sequences of a specific vertebrate virus family by subtraction of the human transcriptisome. PUBLIC HEALTH RELEVANCE: This is an exploratory proposal to determine the viral cause of a chronic central nervous system disease that occurs in the Sahka Republic in far northeastern Russia (Siberia). This disease, Vilyuisk encephalomyelitis, has some similarities to multiple sclerosis and appears to be caused by a human Theilovirus. The murine counterpart, Theiler's murine encephalomyelitis virus, provides a highly relevant experimental animal model for multiple sclerosis.