Arthropod exosomes mediate vector-pathogen interactions

项目摘要/摘要 扁虱传播几种病原体，包括导致人类疾病的黄病毒。这个 节肢动物传播黄病毒到脊椎动物宿主的分子决定因素和机制 人们对此了解甚少。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的细胞系，如扁虱，会分泌介导传播的外体 将黄病毒核糖核酸和蛋白质带到人类细胞。我们注意到壁虱传播的兰加特病毒(LGTV)，一种模型 与壁虱传播的脑炎病毒(TBEV)关系密切的病原体，大量利用壁虱外切体进行 病毒RNA和蛋白质向人皮肤角质形成细胞和血液内皮细胞的传播。冷藏- EM分析显示存在直径在30~200 nm之间的扁虱外切体。 检测阳性和阴性LGTV RNA链和蛋白质，如包膜(E)和非 节肢动物外切体内的结构1(NS1)证实硬虱外切体含有病毒RNA和 蛋白质。来自扁虱和哺乳动物细胞的外切体中的病毒RNA和蛋白质得到了高度保护 在所有经过测试的评估中都具有传染性和复制性。此外，GW4869，a对硬虱细胞的处理。 阻断外体产生的选择性抑制剂影响节肢动物的LGTV载量和传播 对人类细胞来说。一维凝胶电泳法进一步揭示了几种节肢动物富集外切体的存在 扁虱分子。这些初步的结果构成了提出描述角色的强有力的基础 壁虱-LGTV相互作用中的节肢动物外体。描述分子信号和信号的几种方法 提出了节肢动物外体蛋白在Tick-LGTV相互作用中的鉴定方法。我们假设 壁虱外体富集蛋白可被认为是开发抗扁桃体抗体的理想候选蛋白。 媒介疫苗。提出的目标为定义分子基础提供了重要的见解。 硬蜱外切体与病原体的关系。这是一项变革性的新研究，而不是 不仅提供了节肢动物外体在病媒-病原体相互作用中的作用的信息，而且还导致了 发展更好的策略来治疗或控制病原体的传播 其他医学上重要的媒介。

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.