Mechanisms of Chikungunya Virus Binding and Entry

基孔肯雅病毒结合和进入的机制

• 批准号: 8489776
• 负责人: Laurie A. Silva
• 金额: $ 5.22万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8489776
• 关键词: Ablation; Acute; Affect; Africa; Alkanesulfonates; Alphavirus; Antiviral Agents; Antiviral Therapy; Arboviruses; Area; Arthralgia; Arthritis; Asia; Binding; Biological; Biological Assay; Biology; Bite; Carbohydrates; Cell Communication; Cell Line; Cell Surface Receptors; Cell surface; Cell-Matrix Junction; Cells; Chemicals; Chikungunya virus; Chinese Hamster Ovary Cell; Chronic; Clinical; Country; Culicidae; Cultured Cells; Defect; Development; Disease Outbreaks; Enzymes; Epidemic; Exanthema; Fever; Fostering; Foundations; Functional disorder; Genomics; Glycoproteins; Glycosaminoglycans; Health; Heparitin Sulfate; High Prevalence; Human; In Vitro; India; Indian Ocean; Insect Vectors; Integration Host Factors; Invaded; Knowledge; Lead; Licensing; Ligands; Link; Mammalian Cell; Mediating; Membrane Glycoproteins; Molecular; Outcome; Pathogenesis; Play; Point Mutation; Polyarthralgias; Polysaccharides; Prevalence; Proteins; Proteomics; Reporting; Research; Research Personnel; Rheumatism; Role; Screening procedure; Site; Southeastern Asia; Staging; Structure; Techniques; Testing; Therapeutic Intervention; Tropism; United States; Vaccines; Vertebrates; Viral; Viral Envelope Proteins; Viral Fusion Proteins; Viral Pathogenesis; Virulence; Virus; Virus Diseases; Virus Replication; Work; attenuation; base; carbohydrate receptor; cell type; env Gene Products; expression cloning; gain of function; glycosylation; insight; loss of function; mutant; pathogen; programs; receptor; research study; tissue tropism; transmission process; uptake; vector; vector mosquito; virus host interaction; virus tropism

DESCRIPTION (provided by applicant): Chikungunya virus (CHIKV) is an emerging mosquito-borne alphavirus responsible for recent major outbreaks of polyarthralgia in countries of the Indian Ocean region. This important human pathogen causes an acute febrile illness characterized by a maculopapular rash and incapacitating arthralgia, which can evolve to chronic arthritis. CHIKV, like other alphaviruses, is transmitted through the bite of an infected mosquito. The virus is currently endemic in Africa, India, and Southeast Asia, but it has recently spread to new areas with attack rates as high as 75 percent. Due to the high prevalence of the CHIKV insect vectors in many parts of the world, the potential for CHIKV to cause significant epidemics in uninfected regions remains high. Little is known about how this virus interacts with the host and the factors that govern tissue tropism. Receptor engagement plays an essential role in viral replication and thus influences the course and outcome of viral disease. To understand the host range, tissue tropism, and virulence of this pathogen, it is critical to elucidate molecular mechanisms of CHIKV attachment and cell entry. Identifying the attachment molecules to which CHIKV binds is a critical step in obtaining a more comprehensive analysis of the envelope-receptor interactions in CHIKV pathobiology. In addition, understanding key determinants of receptor utilization may provide insight into the factors that facilitate transmission between vector and host. This application investigates the viral and cellular determinants of CHIKV cell entry and will further an understanding of mechanisms by which CHIKV enters host cells. In Specific Aim 1, we will determine the role of N-linked glycosylation sites in the viral envelope proteins in viral replication. Wild-type and glycosylation-site mutant viruses will be produced in mammalian and mosquito cells. The utilization and type of glycosylation at each site will be determined. Cell binding, infectivity, and replication of mutant viruses will be assessed. In Specific Aim 2, the role of cell-surface glycans in CHIKV replication will be investigated. Types of glycans to which CHIKV binds will be identified by glycan array, and their role in virus binding and cell entry will be analyzed. In Specific Aim 3, host proteins that mediate viral entry will be identified using proteomic and genomic strategies. Candidate receptors will be analyzed for their role in facilitating virus infection. These studies will provide valuable insights into mechanisms of CHIKV cell entry and further elucidate virus-host interactions at a critical stage of viral replication. Moreover, findings from these studies will provide new insights into the tissue tropism and pathophysiology of CHIKV, which may illuminate new strategies for therapeutic intervention.

描述（由申请人提供）：Chikungunya病毒（Chikv）是新兴的蚊子 - 传播α病毒，负责在印度洋地区最近发生的多拉尔乳糖爆发。这种重要的人类病原体引起的急性发热疾病，其特征是大绝语皮疹和无行为能力的关节痛，可以演变为慢性关节炎。与其他α病毒一样，Chikv通过被感染的蚊子的咬伤传播。该病毒目前在非洲，印度和东南亚是地方性的，但最近已扩散到攻击率高达75％的新地区。由于世界许多地方Chikv昆虫向量的高流行率，因此Chikv在未感染的地区引起大量流行病的可能性仍然很高。关于该病毒如何与宿主相互作用以及控制组织偏向主义的因素，知之甚少。受体参与在病毒复制中起着至关重要的作用，从而影响病毒疾病的病程和结果。为了了解这种病原体的宿主范围，组织的偏向和毒力，阐明Chikv附着和细胞进入的分子机制至关重要。识别ChiKV结合的依恋分子是对ChiKV病理生物学中的包膜受体相互作用进行更全面分析的关键步骤。此外，了解受体利用的关键决定因素可能会深入了解促进向量和宿主之间传播的因素。该应用研究了Chikv细胞进入的病毒和细胞决定因素，并将进一步了解Chikv进入宿主细胞的机制。在特定的目标1中，我们将确定N连接的糖基化位点在病毒复制中病毒包膜蛋白中的作用。哺乳动物和蚊子细胞将产生野生型和糖基化位点突变病毒。将确定每个位点的糖基化的利用和类型。将评估突变病毒的细胞结合，感染性和复制。在特定的目标2中，将研究细胞表面聚糖在CHIKV复制中的作用。 CHIKV结合的聚糖类型将通过聚糖阵列鉴定，并且将分析它们在病毒结合和细胞进入中的作用。在特定目标3中，将使用蛋白质组学和基因组策略来鉴定介导病毒进入的宿主蛋白。将分析候选受体在促进病毒感染中的作用。这些研究将提供对Chikv细胞进入机制的有价值的见解，并在病毒复制的关键阶段进一步阐明病毒宿主相互作用。此外，这些研究的发现将为CHIKV的组织疗法和病理生理学提供新的见解，这可能会阐明治疗干预的新策略。