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：蜱传播细菌、病毒和寄生虫，导致人类和其他动物疾病。 动物这种现象部分是由于分泌的多余和多能唾液蛋白， 破坏宿主体内平衡并改变血液喂养后的炎症。最近，细胞外囊泡， 介导种间通讯的纳米囊泡的异质群体被证明有助于 病原体传播给哺乳动物我们开发了一种蜱唾液类器官系统， 囊泡释放。我们还操纵了蜱细胞外囊泡的生物合成， N-乙基马来酰亚胺敏感因子附着受体（SNARE）基因的RNA干扰。最后， 我们提供了因果关系，我们的研究结果表明，蜱细胞外囊泡影响居民树突状细胞， 表皮T细胞（DETC）。皮肤中的蜱纳米囊泡如何靶向免疫细胞仍然存在 不安蜱类细胞外囊泡在种间关系中是否具有功能可塑性 仍然难以捉摸在这个R21应用程序中，我们将探讨蜱细胞外囊泡的中心假设， 通过DETC-角质形成细胞为肩胛硬蜱的摄食提供有利的皮肤免疫环境， 轴线DETC与角质形成细胞相互作用，角质形成细胞占皮肤表皮层的约95%。在Aim中 #1这个建议，我们将确定是否影响我。肩胛肌细胞外囊泡对DETC是 直接或间接。我们将使用单细胞RNA测序结合缺乏DETC的动物模型， 评估蜱虫叮咬期间皮肤免疫反应的方向性。在本补助金提案的目标#2中，我们 将确定当蜱细胞外囊泡的生物发生被破坏时皮肤的结构。我们将 通过流式细胞术结合细胞分选通过皮肤活检测量DETC的免疫应答， 微生物刺激和空间转录组学。空间转录组学结合了传统的组织学 通过单细胞基因表达分析和位置条形码化提供信息。因此，我们将构建一个 蜱叮咬期间与DETC和角质形成细胞相关的基因的结构图。总的来说，R21 该项目将强调蜱虫作为节肢动物的重要性，