The role of innate immune cell polarity in the canine respiratory tract following canine distemper virus infection using in vitro and ex vivo models

使用体外和离体模型研究犬瘟热病毒感染后犬呼吸道中先天免疫细胞极性的作用

• 批准号: 408132262
• 负责人: Professor Dr. Wolfgang Baumgärtner, Ph.D.
• 金额: --
• 依托单位: Institut für Pathologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/408132262?language=en
• 关键词: role; innate; immune; cell; polarity

Canine distemper virus (CDV) is a highly contagious morbillivirus, which causes severe systemic disease with involvement of the respiratory tract in domestic and wildlife carnivores. Innate immune cells play a key role in the pathogenesis in a variety of viral respiratory diseases. However, the knowledge about pulmonary innate immunity in canine distemper is still fragmentary. The envisioned project is based on our previous work, in which it could be demonstrated that innate immune cells are able to carry CDV to facilitate cell-to-cell transmission in the respiratory tract and that restriction of antiviral signaling pathways of innate immune cells enhance virus release from the lung in canine distemper. The first part of the project aims at investigating the polarizing effect of CDV upon innate immune cells in vitro. Here the ability of CDV to influence the phenotype of pulmonary and blood-derived macrophages, and the maturation state of monocyte-derived dendritic cells will be analyzed by flow cytometry. In addition, virus-mediated effects upon macrophages and dendritic cells will be characterized by whole transcriptome analyses and functional assays, including mixed leukocyte reaction, as well as migration, phagocytosis and nitric oxide release assays. In the second part, the impact of macrophage polarity and dendritic cell maturation upon CDV cell entry and the capacity of modulated innate immune cells to transmit CDV to the airway epithelium will be investigated using canine air-liquid interface cultures and precision-cut lung slices. In addition, virus-induced cytopathic effects and ultrastructural changes, such as ciliary pathology and apoptosis induction, as well as the regenerative capacity of infected cultures will be determined. The study will give mechanistic insights in the dysfunction of pulmonary innate immunity in CDV infection and its impact on disease pathogenesis. Elucidating the regulatory mechanisms through which pathogens regulate innate immune cell plasticity will contribute to the discovery of therapeutic targets in morbillivirus diseases and prevention of virus transmission to other hosts.

犬瘟热病毒（CDV）是一种高度传染性的麻疹病毒，可引起家畜和野生食肉动物严重的呼吸道系统疾病。天然免疫细胞在多种病毒性呼吸道疾病的发病机制中起关键作用。然而，对犬瘟热肺部天然免疫的认识仍然是零碎的。设想的项目是基于我们以前的工作，其中可以证明先天免疫细胞能够携带CDV以促进呼吸道中的细胞间传播，并且先天免疫细胞的抗病毒信号传导途径的限制增强犬瘟热中病毒从肺部释放。第一部分研究CDV对先天性免疫细胞的极化作用。在此，将通过流式细胞术分析CDV影响肺和血液来源的巨噬细胞的表型以及单核细胞来源的树突状细胞的成熟状态的能力。此外，病毒介导的对巨噬细胞和树突状细胞的影响将通过全转录组分析和功能测定来表征，包括混合白细胞反应以及迁移、吞噬作用和一氧化氮释放测定。在第二部分中，巨噬细胞极性和树突状细胞成熟后CDV细胞进入和调制的先天免疫细胞的能力，以传输CDV气道上皮细胞的影响将使用犬气液界面培养和精密切割肺切片进行研究。此外，还将确定病毒诱导的细胞病变效应和超微结构变化，如纤毛病理学和细胞凋亡诱导，以及感染培养物的再生能力。该研究将为CDV感染中肺部先天免疫功能障碍及其对疾病发病机制的影响提供机制见解。阐明病原体调节先天免疫细胞可塑性的调节机制将有助于发现麻疹病毒疾病的治疗靶点和预防病毒传播到其他宿主。