Characterization of alphavirus infection receptors

描述（由申请人提供）：甲病毒是节肢动物载体病毒，可引起从轻度发热性疾病到严重关节痛或致命脑炎的人类疾病。由于一些因素，包括易于隔离、长期稳定和以气雾剂施用时可能具有高发病率/死亡率，其中一些已被指定为可能用于生物战/生物恐怖主义的B类选定制剂。不幸的是，目前还没有获得许可的疫苗或抗病毒药物来对抗这些感染。阻断病毒与细胞的初始相互作用是预防感染和疾病的最有效手段之一，了解病毒与感染受体的相互作用对设计感染阻断药物至关重要。然而，对于尚未广泛适应细胞培养生长的甲病毒的受体相互作用知之甚少。利用一种新的载体系统，我们提出同时检测多种具有代表性的新世界和旧世界菌株的非细胞适应甲型病毒的受体相互作用。在第一个目标中，我们将研究利用先前表征的细胞表面受体分子硫酸乙酰肝素和高亲和力层粘胶蛋白受体，使用表征良好的细胞系。在第二个目标中，我们将利用我们的新载体系统和cDNA文库筛选中最近鉴定的α病毒受体阴性细胞系来鉴定可以介导细胞α病毒感染的新分子。这些实验结果将提供对新旧大陆甲病毒受体相互作用的全面了解，并为设计感染阻断抗病毒药物提供框架。公共卫生相关性：我们的计划是使用一种新的嵌合甲型病毒复制子系统，其中将一种繁殖缺陷的委内瑞拉马脑炎病毒基因组包装在不同的旧大陆和新世界甲型病毒的结构蛋白中，以比较这些病毒低传代株的受体利用特征。这些比较将包括直接评估先前鉴定的受体（硫酸肝素，高亲和力层粘连蛋白受体）介导感染的能力，以及通过使用逆转录病毒表达文库和甲病毒受体阴性细胞表征新受体。

DESCRIPTION (provided by applicant): Alphaviruses are arthropod-vectored viruses that can cause human diseases ranging from mild febrile illness to severe arthralgia or fatal encephalitis. Due to several factors including the ease of their isolation, long-term stability and potential for high morbidity/mortality when administered by aerosol, several have been designated Category B Select Agents with potential for biowarfare/bioterrorism use. Unfortunately, no licensed vaccines or antiviral medications are available to combat these infections. Interruption of the initial interactions of viruses with cells is one of the most effective means of preventing infection and disease and an understanding of virus-infection receptor interactions is vital to design of infection-blocking drugs. However, little is known about the receptor interactions of alphaviruses that have not been adapted extensively to growth in cell culture. Using a novel vector system, we propose to examine simultaneously, the receptor interactions of multiple non-cell adapted alphaviruses representative of both New World and Old World strains. In the first Aim, we will investigate the utilization of previously characterized cell surface receptor molecules heparan sulfate and the high affinity laminin receptor using well-characterized cell lines. In the second Aim we will utilize our novel vector system and a recently-identified alphavirus receptor-negative cell line in a cDNA library screen to identify new molecules that can mediate alphavirus infection of cells. The results of these experiments will provide a comprehensive understanding of the receptor interactions of New World and Old World alphaviruses and provide a framework for design of infection-blocking antiviral drugs. PUBLIC HEALTH RELEVANCE: Our plan is to use a novel chimeric alphavirus replicon system in which a propagation-defective Venezuelan equine encephalitis virus genome is packaged in the structural proteins of different Old World and New World alphaviruses to compare the receptor utilization characteristics of low passage strains of these viruses. These comparisons will include direct evaluation of the capacity of previously-identified receptors (heparan sulfate, the high affinity laminin receptor) to mediate infection as well characterization of new receptors through use of a retrovirus expression library and alphavirus receptor-negative cells.