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)是布尼亚病毒科奈罗病毒属的成员，可引起人类严重疾病，死亡率高(50%)。人类可通过被硬蜱叮咬、在感染急性期接触CCHFV感染者或接触病毒型家畜的血液或组织而感染。基于CCHFV的高死亡率和CCHFV通过气雾化传播的潜力，CCHFV被认为具有潜在的生物恐怖主义重要性，并被归类为B类病原体。由于要求在BSL4条件下与CCHFV一起工作，人们几乎对其细胞生物学、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。猪瘟病毒基因组由3个负义片段组成：小片段(S)、中片段(M)和大片段(L)。CCHFV RNA M片段编码病毒糖蛋白G1和G2。G1蛋白的不同寻常之处在于，它有一个N-末端的粘蛋白样结构域，该结构域是在翻译后从蛋白质中切割出来的，第二个N-末端结构域也是从蛋白质中切割出来的，我们最近发现，也会发生一个C-末端的切割事件，导致蛋白质的大部分细胞质区域的释放。在过去的一年里，我们与康妮·施马尔约翰博士(USAMRIID)和Adolfo Garcia-Sastre博士(西奈先生)共同发起了一项关于CCHFV的合作研究。我们的长期计划是准备一项计划项目赠款来支持我们的工作。此R21应用程序的目的是为DOMS实验室的研究寻求支持，该实验室将提供P01应用程序所需的初步结果。在R21的这一应用中，我们提出了以下三个具体目标：目的1.从不同地理位置的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。目的2.鉴定一组新的针对CCHFV G1和G2蛋白的单抗，包括一些中和抗体。目的3.建立病毒假型系统或细胞-细胞融合试验，用于研究CCHFV在BSL2或BSL3条件下的趋向性、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.