Crimean congo hemorrhagic fever virus glycoproteins

描述（由申请人提供）： 克里米亚-刚果出血热病毒（CCHFV）是布尼亚病毒科Nairovirus属的成员，在人类中引起严重疾病，具有高死亡率（50%）。人类可通过蜱叮咬、在感染急性期与CCHFV感染患者接触或与病毒血症牲畜的血液或组织接触而感染。基于CCHFV的高死亡率和CCHFV通过雾化传播的可能性，CCHFV被认为具有潜在的生物恐怖主义重要性，并被分类为B类制剂。由于需要在BSL 4条件下与CCHFV一起工作，实际上对其细胞生物学、其进入细胞的机制、其糖蛋白的抗原结构或抗体可以中和病毒的机制一无所知。CCHFV的RNA基因组由三个负义片段组成：小（S）、中（M）和大（L）基因组RNA片段。CCHFV RNA M片段编码病毒糖蛋白G1和G2。G1蛋白是不寻常的，因为它有一个N-末端粘蛋白样结构域，从蛋白质裂解后，第二个N-末端结构域，也从蛋白质裂解，我们最近发现，C-末端裂解事件发生，以及，导致释放的蛋白质的胞质结构域的大部分。在过去的一年里，我们与Connie Schmaljohn博士（USAMRIID）和Adolfo Garcia-Sastre博士（Sinai先生）开展了CCHFV的合作研究。我们的长期计划是准备一个计划项目赠款，以支持我们的工作。本R21申请的目的是为Doms实验室的研究寻求支持，这些研究将提供P01申请所需的初步结果。在这个R21应用中，我们提出了以下三个具体目标：目标1。从地理上不同的CCHFV分离株中克隆和分析CCHFV M蛋白片段，并开发研究和比较G1和G2蛋白所需的表达系统和工具。目标2.鉴定一组新的针对CCHFV G1和G2蛋白的单克隆抗体，包括一些中和抗体。目标3.开发病毒假型系统或细胞-细胞融合试验，可用于研究CCHFV在BSL 2或BSL 3条件下的嗜性、融合和进入机制。

DESCRIPTION (provided by applicant): Crimean-Congo Hemorrhagic Fever virus (CCHFV), a member of the genus Nairovirus of the family Bunyaviridae, causes severe disease in humans with high rates of mortality (50%). Humans can become infected through bites of the Ixodid tick, by contact with a CCHFV infected patient during the acute phase of infection, or by contact with blood or tissue from viremic livestock. Based on the high mortality rate of CCHFV and the potential for CCHFV dissemination by aerosolization, CCHFV is considered to be of potential bioterrorism importance and is classified as a Category B agent. Because of the requirement to work with CCHFV under BSL4 conditions, virtually nothing is known about its cell biology, the mechanisms by which it enters cells, the antigenic structure of its glycoproteins or the mechanisms by which antibodies can neutralize the virus. The RNA genome of CCHFV consists of three negative sense segments: the small (S), medium (M) and large (L) genomic RNA segments. The CCHFV RNA M segment encodes the virus glycoproteins, G1 and G2. The G1 protein is unusual in that it has an N-terminal mucin-like domain that is cleaved from the protein post-translationally, a second N-terminal domain that is also cleaved from the protein, and we have recently found that a C-terminal cleavage event occurs as well, resulting in release of much of the cytoplasmic domain of the protein. Over the past year, we have initiated a collaborative study of CCHFV with Dr. Connie Schmaljohn (USAMRIID) and Dr. Adolfo Garcia-Sastre (Mr. Sinai). Our long-term plan is to prepare a Program Project grant to support our work. The intent of this R21 application is to seek support for studies in the Doms lab that will provide the preliminary results needed for the P01 application. In this R21 application, we propose the following three Specific Aims: Aim 1. Clone and analyze CCHFV M protein segments from geographically diverse CCHFV isolates, and develop the expression systems and tools needed to study and compare the G1 and G2 proteins. Aim 2. Characterize a novel panel of monoclonal antibodies directed against the CCHFV G1 and G2 proteins, including a number of neutralizing antibodies. Aim 3. Develop virus pseudotype systems or cell-cell fusion assays that can be used to study CCHFV tropism, fusion and entry mechanisms under BSL2 or BSL3 conditions.