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Arthropod exosomes mediate vector-pathogen interactions

节肢动物外泌体介导载体-病原体相互作用

基本信息

批准号：
10668352
负责人：
Hameeda Sultana
金额：
$ 36.5万
依托单位：
UNIVERSITY OF TENNESSEE KNOXVILLE
依托单位国家：
美国
项目类别：
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/10668352
关键词：
Address Affect Area Arthropod Vectors Arthropods Biochemical Biological Biological Assay Biology Black-legged Tick Blood Cell Line Cells Data Detection Development Diameter Disease Electron Microscopy Electrons Endothelial Cells Enzymes Flavivirus Flavivirus Infections Gel Genes Goals Human Immune response Immunization In Vitro Infection Langat virus Lead Life Cycle Stages Mammalian Cell Mediating Mediator Medical Microscopic Migration Assay Modeling Molecular Mus Paper Production Proteins Proteomics Publishing RNA RNA Interference Research Role Salivary Gland Tissue Salivary Glands Secure Signal Transduction Skin Sphingomyelinase Techniques Testing Tick-Borne Encephalitis Virus Ticks Time Two-Dimensional Gel Electrophoresis Validation Vector-transmitted infectious disease Vesicle Viral Viral Proteins Viral load measurement Virulent Virus Virus Diseases arboviral disease arthropod-borne cell type evaluation/testing exosome extracellular feeding gel electrophoresis inhibitor insight keratinocyte knock-down late endosome microvesicles mouse model novel pathogen recruit therapeutic evaluation tick feeding tick saliva tick transmission tick-borne tick-borne flavivirus transmission process transmission-blocking vaccine vector vector tick vector transmission vector vaccine viral RNA viral transmission

项目摘要

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.

项目总结/摘要蜱传播几种病原体，包括引起人类疾病的黄病毒。的节肢动物传播黄病毒的分子决定因素和机制我们对此知之甚少。在这项研究中，我们提供了强有力的初步数据，首次表明，来自医学上重要的节肢动物如蜱的细胞系分泌介导传播的外泌体将黄病毒RNA和蛋白质导入人体细胞。我们注意到，蜱传Langat病毒（LGTV），一个模型，与蜱传脑炎病毒（TBEV）密切相关的病原体，大量使用蜱外泌体，将病毒RNA和蛋白质传输到人体皮肤角质细胞和血液内皮细胞。冷冻- EM分析显示存在直径为30至200 nm的大小范围的蜱外泌体。检测阳性和阴性LGTV RNA链和蛋白质，如包膜（E）和非包膜蛋白（Non- 节肢动物外泌体内部结构1（NS 1）证实蜱外泌体含有病毒RNA， proteins.来自蜱和哺乳动物细胞的外泌体中的病毒RNA和蛋白质被高度安全地固定，感染性和复制性。此外，用GW 4869（a 阻断外泌体产生的选择性抑制剂影响LGTV负载和从节肢动物的传播到人体细胞。1-D凝胶电泳进一步揭示了几种节肢动物外泌体富集的存在。蜱分子。这些初步结果构成了强有力的基础，为建议的特点作用，节肢动物外来体在蜱LGTV相互作用。几种描述分子信号传导的方法，提出了在蜱-LGTV相互作用中识别节肢动物外泌体蛋白的方法。我们假设蜱外泌体富集蛋白可以被认为是开发抗- 载体疫苗。提出的目标提供了重要的见解，以确定分子基础的蜱外泌体和病原体之间的关系。这是一项变革性的新颖研究，不仅提供了节肢动物外泌体在媒介-病原体相互作用中的作用的信息，发展更好的策略来治疗或控制病原体的传播，其他重要的医学载体。