Regulation of Skin Immunity by a Tick Bite

蜱虫叮咬对皮肤免疫的调节

• 批准号: 10514626
• 负责人: Joao Pedra
• 金额: $ 19.31万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10514626
• 关键词: Affect; Anesthetics; Animal Model; Animals; Anti-Inflammatory Agents; Anticoagulants; Applications Grants; Arboviruses; Architecture; Area; Arthropods; Bacteria; Bar Codes; Biogenesis; Biological Assay; Biology; Biopsy; Bite; Black-legged Tick; Blood; Castor Bean Tick; Cattle; Cavia; Cell Communication; Cell Separation; Cells; Chromosome Mapping; Communities; Coupled; Dermal; Dermis; Disease; Emerging Technologies; Environment; Epidermis; Etiology; Europe; Fermentation; Flow Cytometry; Future; Gene Expression Profiling; Hemostatic Agents; Histology; Homeostasis; Human; Immune; Immune response; Immunity; Immunobiology; Immunology procedure; Inflammation; Intervention; Knowledge; Laboratories; Literature; Mammals; Measures; Mediating; Molecular; Molecular Analysis; Mus; N-ethylmaleimide-sensitive protein; Organoids; Parasites; Population; Property; Proteins; RNA; RNA Interference; Reagent; Receptor Gene; Regulation; Research; Research Personnel; Resistance; Resources; Role; Saliva; Salivary; Salivary Glands; Salivary Proteins; Seminal; Site; Skin; System; T-Lymphocyte; Technology; Therapeutic; Thinking; Tick-Borne Diseases; Ticks; Travel; United States; Virus; Work; acquired immunity; arthropod-borne; extracellular vesicles; fascinate; feeding; follow-up; foot; functional plasticity; immunoregulation; insight; interspecies communication; keratinocyte; microbial; nanovesicle; neglect; new technology; pathogen; receptor; single-cell RNA sequencing; tick bite; tick feeding; tick saliva; tick transmission; tool; transcriptomics; transmission process; vector; vesicular release; wound healing; γδ T cells

Summary/Abstract: Ticks transmit bacteria, viruses and parasites that cause disease in humans and other animals. This phenomenon is partly due to the secretion of redundant and pluripotent salivary proteins that disrupt host homeostasis and alter inflammation upon blood-feeding. Recently, extracellular vesicles, a heterogenous population of nanovesicles that mediate interspecies communication, were shown to facilitate pathogen transmission to mammals. We developed a tick salivary organoid system that mimics extracellular vesicle release. We also manipulated the biogenesis of tick extracellular vesicles by silencing the expression of N-ethylmaleimide-sensitive factor attachment receptor (SNARE) genes through RNA interference. Finally, we provided causality to our findings by showing that tick extracellular vesicles affect resident dendritic epidermal T cells (DETCs) in the skin. How immune cells are targeted by tick nanovesicles in the skin remains unsettled. Whether tick extracellular vesicles have functional plasticity during interspecies relationships remains elusive. In this R21 application, we will explore the central hypothesis that tick extracellular vesicles provide an advantageous skin immune environment for Ixodes scapularis feeding via the DETC-keratinocyte axis. DETCs interact with keratinocytes, which comprise approximately 95% of the skin epidermal layer. In Aim #1 of this proposal, we will determine whether the effect of I. scapularis extracellular vesicles on DETCs is direct or indirect. We will use single cell RNA sequencing coupled with animal models devoid of DETCs to evaluate the directionality of the skin immune response during a tick bite. In Aim #2 of this grant proposal, we will ascertain the architecture of the skin when the biogenesis of tick extracellular vesicles is disrupted. We will measure the immune response of DETCs through skin biopsies via flow cytometry coupled with cell sorting, microbial stimulation and spatial transcriptomics. Spatial transcriptomics combines traditional histology information with single cell gene expression analysis and positional barcoding. Thus, we will construct an architectural map of genes associated with DETCs and keratinocytes during a tick bite. Collectively, this R21 project will underscore the importance of ticks as arthropods of

摘要/摘要：扁虱传播细菌、病毒和寄生虫，这些细菌、病毒和寄生虫会导致人类和其他疾病 动物。这种现象的部分原因是分泌了多余的和多能的唾液蛋白 破坏宿主体内平衡，改变血液摄食时的炎症。最近，细胞外小泡，一种 异质的纳米卵泡种群，调解物种间的交流，被证明有助于 病原体传播给哺乳动物。我们开发了一种模仿细胞外的扁虱唾液有机系统。 囊泡释放。我们还通过沉默表达来操纵壁虱细胞外囊泡的生物发生。 通过RNA干扰获得N-乙基马来酰亚胺敏感因子附着受体(SNARE)基因。最后， 我们提供了我们的发现的因果关系，通过显示壁虱细胞外小泡影响常驻树突状细胞。 皮肤中的表皮T细胞(DETCs)。皮肤中的扁虱纳米囊如何将免疫细胞作为靶点仍然存在 动荡不安。壁虱胞外小泡在种间关系中是否具有功能可塑性 仍然难以捉摸。在R21的应用中，我们将探索勾选细胞外小泡的中心假设 通过DETC角质形成细胞为肩部硬蜱的摄食提供有利的皮肤免疫环境 轴心。DETCs与角质形成细胞相互作用，角质形成细胞约占皮肤表皮的95%。在AIM 在这项提案中，我们将确定肩胛肌细胞外小泡对DETCs的影响是否 直接或间接。我们将使用单细胞RNA测序结合缺乏DETCs的动物模型来 评估扁虱叮咬时皮肤免疫反应的方向性。在这项赠款提案的目标2中，我们 当壁虱细胞外小泡的生物发生被破坏时，将确定皮肤的结构。我们会 流式细胞仪结合细胞分选通过皮肤活检检测DETCs的免疫应答， 微生物刺激和空间转录。空间转录学结合传统组织学 单细胞基因表达分析和位置条形码的信息。因此，我们将构建一个 扁虱叮咬期间与DETCs和角质形成细胞相关的基因架构图。总的来说，这款R21 该项目将强调扁虱作为节肢动物的重要性