Filovirus/cellular receptor interactions

描述（由申请方提供）：丝状病毒在非洲引起周期性毁灭性出血热暴发。由于这些感染引起的高死亡率和这些病毒在人群中的高传播性，这组病毒已被置于疾病控制和预防中心建立的A类选择代理列表中。几年前，埃博拉病毒和马尔堡病毒进入哺乳动物细胞的细胞受体被确定为叶酸受体。随后的研究表明，叶酸受体-a不作为气道上皮或巨噬细胞中的埃博拉受体，这些研究提供的证据表明，叶酸受体-a在丝状病毒进入中的作用一般受到质疑。在这里，我们将使用两种独立的方法来鉴定丝状病毒的细胞受体：1）基于生物信息学的方法和2）功能性丝状病毒糖蛋白结合方法。在第一种方法中，将丝状病毒糖蛋白假型化逆转录病毒构建体转导到一系列充分表征的人肿瘤系（NCI 60细胞）中。将病毒转导数据与这些细胞系的基因表达谱相关联，以鉴定跨膜结构域和GPI连接的蛋白作为丝状病毒受体的潜在候选物。在使用这些细胞系的初步研究中，一种新的跨膜结构域蛋白已被鉴定为候选受体。同时，将通过亲和纯化方法纯化潜在的病毒受体。将在病毒体覆盖蛋白结合试验（VOPBA）中检测所得纯化级分与丝状病毒糖蛋白结合的能力。然后将进行研究，以确定候选受体是否是埃博拉和马尔堡糖蛋白依赖性进入的必要和充分条件。最后，将确定支持丝状病毒感染的潜在受体的功效。丝状病毒用于体内感染的细胞受体的鉴定是开发针对这些病毒的抗病毒剂的重要步骤。对HIV的细胞辅助受体的鉴定导致了阻断辅助受体使用的新药的开发。开发针对丝状病毒的等效药物可能会成功限制新的疫情。

DESCRIPTION (provided by applicant): Filoviruses cause periodic devastating hemorrhagic fever outbreaks in Africa. Because of the high rates of mortality caused by these infections and high transmissibility of these viruses in the human population, this group of viruses has been placed on the Category A select agent list that has been established by the Centers for Disease Control and Prevention. The cellular receptor for Ebola and Marburg virus entry into mammalian cells was identified as folate receptor-a several years ago. Subsequent studies have indicate that folate receptor-a does not serve as the Ebola receptor in airway epithelium or macrophages and these investigations provide evidence that calls the role of the folate receptor-a in entry of filoviruses into question in general. Here we will use two independent approaches to identify cellular receptor(s) for filoviruses: 1) a bioinformatics-based approach and 2) a functional filovirus glycoprotein binding approach. In the first approach, filoviral glycoprotein pseudotyped retroviral constructs will be transduced into a series of well- characterized human tumor lines (NCI60 cells). Viral transduction data will be correlated with the gene expression profiles for these lines to identify transmembrane domain and GPI-linked proteins as potential candidates for filovirus receptors. In preliminary studies using these cell lines, one novel transmembrane domain protein has been identified as a candidate receptor. In parallel, potential viral receptors will be purified through affinity purification approaches. Resulting purified fractions will be tested for their ability to bind to filovirus glycoproteins in virion overlay protein binding assays (VOPBAs). Studies will then be performed to determine if the candidate receptor(s) are necessary and sufficient for Ebola and Marburg glycoprotein dependent entry. Finally, the efficacy of the potential receptor to support filovirus infection will be determined. Identification of cellular receptor(s) that are utilized by filoviruses for in vivo infection is an important step in developing anti-viral agents to these viruses. Identification of the cellular co-receptors for HIV resulted in the development of new drugs that block co-receptor usage. Development of equivalent drugs for the filoviruses might prove to be successful in limiting new outbreaks.