Arthropod exosomes mediate vector-pathogen interactions

节肢动物外泌体介导载体-病原体相互作用

• 批准号: 10322352
• 负责人: Hameeda Sultana
• 金额: $ 35.98万
• 依托单位: UNIVERSITY OF TENNESSEE KNOXVILLE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-19 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322352
• 关键词: Arthropod; exosomes; mediate; vector; pathogen

Project Summary/Abstract Ticks transmit several pathogens including flaviviruses that cause diseases in humans. The molecular determinants and mechanisms of arthropod-borne flavivirus transmission to the vertebrate host are poorly understood. In this study, we provide strong preliminary data that show for the first time that a cell line from medically important arthropods, such as ticks, secretes exosomes that mediate transmission of flavivirus RNA and proteins to the human cells. We noted that tick-borne Langat virus (LGTV), a model pathogen closely related to tick-borne encephalitis virus (TBEV), profusely uses tick exosomes for transmission of viral RNA and proteins to the human- skin keratinocytes and blood endothelial cells. Cryo- EM analysis revealed presence of tick exosomes with the size range of 30 to 200 nm in diameter. Detection of both positive and negative LGTV RNA strands and proteins such as Envelope (E) and Non- structural 1 (NS1) inside arthropod exosomes confirmed that tick exosomes contain viral RNA and proteins. Viral RNA and proteins in exosomes derived from tick and mammalian cells were secured, highly infectious and replicative in all tested evaluations. Furthermore, treatment of tick cells with GW4869, a selective inhibitor that blocks exosome production affected LGTV loads and transmission from arthropod to human cells. 1-D gel electrophoresis further revealed presence of several arthropod exosome-enriched tick molecules. These preliminary results form the strong basis for the proposal to characterize role of arthropod exosomes in tick-LGTV interactions. Several approaches that delineate molecular signaling and identification of arthropod exosomal proteins in tick-LGTV interactions are proposed. We hypothesize that tick exosomal-enriched proteins could be considered as ideal candidates for the development of anti- vector vaccines. The proposed aims provide important insights to define molecular basis of the relationship between tick exosomes and pathogens. This is a transformative and a novel study that not only provides information on the role of arthropod exosomes in vector-pathogen interactions but also lead to the development of better strategies to treat or control transmission of pathogens from this and perhaps other vectors of medical importance.

项目摘要/摘要 tick传递了几种病原体，包括引起人类疾病的黄病毒。这 节肢动物传播黄病毒传播到脊椎动物宿主的分子决定因素和机制 知之甚少。在这项研究中，我们提供了强大的初步数据，这是首次表明 来自医学上重要的节肢动物（例如tick）的细胞系，分泌介导传播的外泌体 黄病毒RNA和人类细胞的蛋白质。我们注意到Tick-Borne Langat病毒（LGTV），一个模型 病原体与tick传播脑炎病毒（TBEV）密切相关，大量使用tick外泌体进行 病毒RNA和蛋白质传播到人体皮肤角质形成细胞和血液内皮细胞。冷冻 EM分析表明，直径30至200 nm的大小范围的tick外泌体存在。 检测阳性和阴性LGTV RNA链和蛋白质（例如包膜（E）和非 - 结构1（NS1）内部节肢动物外泌体中证实tick外泌体含有病毒RNA和 蛋白质。固定从tick和哺乳动物细胞衍生的外泌体中的病毒RNA和蛋白质，高度 在所有经过测试的评估中具有传染性和复制性。此外，用GW4869的壁虱细胞处理 选择性抑制剂阻断外泌体产生影响LGTV负载和节肢动物的传播 到人类细胞。 1-D凝胶电泳进一步揭示了几种节肢动物外泌体富含的存在 滴答分子。这些初步结果构成了提出表征作用的强大基础 tick-LGTV相互作用中的节肢动物外泌体。几种描述了分子信号传导和 提出了tick-LGTV相互作用中节肢动物外泌体蛋白的鉴定。我们假设这一点 tick外泌尿体富集蛋白可以被视为抗肿瘤发展的理想候选者 载体疫苗。提出的目的提供了重要的见解，以定义分子基础 tick外泌体和病原体之间的关系。这是一项变革性和新颖的研究，不是 仅提供有关节肢动物外泌体在载体 - 病原体相互作用中的作用的信息 制定更好的策略来治疗或控制病原体传播的策略，也许 其他重要性的向量。