Immunity to Live Mosquito Probing and Flavivirus Infection in Human Skin

摘要 节肢动物传播的病毒在全球范围内是对人类的主要威胁，这些黄病毒 由埃及伊蚊传播的蚊子是对人类健康最大的威胁。了解 在人类宿主中发生的免疫事件，特别是在病毒传播的皮肤中 蚊子取食，代表了我们对虫媒病毒疾病认识的一大空白。解决这一差距是一项 筹资机会的中心主题PAR-18-860，“对节肢动物吸血的免疫反应”，以及 是当前提案的目标。为了更好地定义这些事件，我们建议进行一项跨学科研究， Barratt-Boyes实验室，在人类皮肤免疫学方面的专业知识，以及Vasilakis实验室，在 植物病毒学和蚊子-病毒-宿主的相互作用。Barratt-Boyes研究小组已经建立了一个体外模型 利用匿名组织的大面积外植体检测登革热和寨卡病毒在人皮肤中的感染 健康供体，并定义了在没有蚊子的情况下皮肤发生的免疫学事件 探查。瓦西拉基斯集团独一无二地进入获准与活体伊蚊合作的昆虫设施 SPP.感染DENV或ZIKV使我们现在可以探索蚊子对病毒的贡献 发病机制。因此，我们将汇集三个基本组件--活蚊子、人类皮肤和 致病病毒-首次解决病媒传播过程中发生的免疫事件 黄病毒在人体皮肤中的传播。这是一个真正创新的提议；据我们所知，没有 现有的出版物描述了人类皮肤对由活体传播的致病性黄病毒的反应 蚊子，没有其他研究小组有像我们这样的实验系统来填补这一研究空白。 我们有两个具体的目标：(1)人类皮肤对蚊子在 没有病毒，以及(2)蚊子探测如何影响DENV和ZIKV在人类中的复制和传播 皮肤。我们的研究结果将对免疫学和传播学领域产生强大和持久的影响 由病媒传播的病原体。这项工作将建立体外蚊子皮肤病毒系统，作为 未来在发病机制、治疗和疫苗开发方面的研究。

Abstract Arthropod-borne viruses represent a major threat to humans on a global scale, and of these flaviviruses transmitted by Aedes aegypti mosquitoes are the most significant risk to human health. Understanding the immunologic events that take place in the human host, particularly in the skin where virus is transmitted during mosquito feeding, represents a major gap in our understanding of arbovirus disease. Addressing this gap is a central theme of the Funding Opportunity PAR-18-860, “Immune Response to Arthropod Blood Feeding”, and is the goal of the current proposal. To best define these events, we propose a transdisciplinary study between the Barratt-Boyes lab, with expertise in human skin immunology, and the Vasilakis lab, with expertise in arbovirology and mosquito-virus-host interactions. The Barratt-Boyes group has established an ex vivo model of dengue (DENV) and Zika virus (ZIKV) infection in human skin using large-area explants from anonymous healthy donors, and have defined the immunologic events that occur in skin in the absence of mosquito probing. The unique access of the Vasilakis group to insectary facilities approved for working with live Aedes spp. infected with DENV or ZIKV allows us to now explore the contribution of the mosquito to viral pathogenesis. Thus, we will bring together three essential components – live mosquitoes, human skin, and pathogenic viruses – to address for the first time the immunologic events that occur during vector-borne transmission of flaviviruses in human skin. This is a truly innovative proposal; to our knowledge, no publications exist describing the response of human skin to pathogenic flaviviruses transmitted by live mosquitoes, and no other research group has an experimental system such as ours to fill this research gap. We have two specific aims: (1) what is the immune response of human skin to mosquito probing in the absence of virus, and (2) how does mosquito probing affect DENV and ZIKV replication and spread in human skin. The results of our study will have a strong and lasting impact on the field of immunology and transmission of vector-borne pathogens. The work will establish the ex vivo mosquito-skin-virus system as a foundation for future studies on pathogenesis, therapy and vaccine development.