DENGUE VIRUS INTERACTION WITH TARGET CELL RECEPTORS

登革热病毒与靶细胞受体的相互作用

• 批准号: 2852898
• 负责人: RORY M MARKS
• 金额: $ 33.12万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2852898
• 关键词: antiviral agents; dengue virus; drug design /synthesis /production; gene mutation; proteoglycan; recombinant proteins; site directed mutagenesis; suramin; tissue /cell culture; transfection; virus infection mechanism; virus protein; virus receptors

Dengue virus is a human pathogenic flavivirus that has re-emerged as a serious public health threat. There is no treatment or vaccine, and public health control measures have failed. There is little understanding of the pathogenesis of dengue virus infection (or that of any other flavivirus). The binding of viral ectodomain molecules to specific receptors on target cells is a critical factor in pathogenicity as it is an important determinant of cell and tissue tropism; yet, there is no information about the molecular basis of the binding of dengue virus to target cells. We have characterized the interaction of dengue virus envelope protein with target cells, and partially identified a target cell receptor. We propose to fully identify the proteoglycan that is a cellular receptor for dengue virus, assess for the presence of co-receptor mechanisms, identify the envelope protein domains responsible for cell-binding, and develop inhibitors of infection based on blockade of viral target cell binding. Aim 1 will identify the proteoglycan (PG) that is the dengue receptor for Vero and other cells. Results indicate that the glycosaminoglycan (GAG) component of this PG embodies the receptor specificity, and is an unusually highly sulfated form of heparan sulfate. Aim 2 will identify the dengue envelope protein motifs responsible for target cell binding. Results indicate that the envelope protein accounts for viral binding, and has GAG-binding domains in two regions at the carboxy-terminus. Recombinant envelope proteins with mutations in these regions will be expressed, and their binding properties compared with wild-type envelope protein. Infectious virus incorporating mutations of interest will be generated, and assessed for changes in infectivity. Aim 3 will identify pharmaceuticals that prevent infection by blocking virus - target cell binding. Results indicate that the polysulfonate Suramin inhibits both envelope protein binding, and infectivity. Using the structure of Suramin as a guide, Suramin analogues and other sulfated/sialylated molecules will be synthesized, and tested for activity in inhibiting envelope protein binding to, and dengue virus infection of, target cells. Viruses subvert cell-surface molecules involved in mediating physiological processes to gain access to target cells. In addition to understanding this important element of dengue virus pathogenesis, identification of dengue target cell receptors may also lead to an understanding of the physiological roles of these cellular receptors, as well as advances in understanding interactions between complex carbohydrates and proteins.

登革热病毒是一种人类致病性黄病毒，已作为一种病毒重新出现。 严重的公共卫生威胁。 没有治疗方法或疫苗，并且 公共卫生控制措施失败了。 很少有 了解登革热病毒感染（或 任何其他黄病毒）。 病毒胞外域分子与 靶细胞上的特异性受体是致病性的关键因素 因为它是细胞和组织趋向性的重要决定因素；然而，有 没有关于登革热结合的分子基础的信息 病毒到达靶细胞。我们已经描述了登革热的相互作用 病毒包膜蛋白与靶细胞，并部分鉴定出 靶细胞受体。 我们建议充分鉴定蛋白多糖 这是登革热病毒的细胞受体，评估其存在 共同受体机制，识别包膜蛋白结构域 负责细胞结合，并开发基于感染的抑制剂 阻断病毒靶细胞结合。 目标 1 将识别作为登革热受体的蛋白多糖 (PG) 对于 Vero 和其他细胞。 结果表明，糖胺聚糖 该PG的（GAG）成分体现了受体特异性，是一种 硫酸乙酰肝素的异常高度硫酸化形式。 目标 2 将确定 负责靶细胞结合的登革热包膜蛋白基序。 结果表明包膜蛋白负责病毒结合， 并在羧基末端的两个区域具有 GAG 结合域。 这些区域发生突变的重组包膜蛋白将被 表达，以及与野生型包膜相比的结合特性 蛋白质。 包含感兴趣突变的传染性病毒将被 生成并评估传染性的变化。 目标 3 将确定 通过阻断病毒-靶细胞来预防感染的药物 绑定。 结果表明，多磺酸盐苏拉明同时抑制 包膜蛋白结合和感染性。 使用结构 苏拉明作为指导，苏拉明类似物和其他硫酸化/唾液酸化 将合成分子并测试其抑制活性 包膜蛋白与靶标结合以及登革热病毒感染靶标 细胞。 病毒破坏参与介导的细胞表面分子 进入靶细胞的生理过程。 此外 了解登革热病毒发病机制的这一重要因素， 登革热靶细胞受体的鉴定也可能导致 了解这些细胞受体的生理作用， 以及理解复杂之间相互作用的进展 碳水化合物和蛋白质。