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.

犬科病毒（CDV）是一种高度具有传染性的虫害病毒，引起严重的全身性疾病，呼吸道参与家庭和野生动植物食肉动物。先天免疫细胞在多种病毒呼吸道疾病的发病机理中起关键作用。但是，关于犬科动物中肺先天免疫的知识仍然是零散的。设想的项目基于我们以前的工作，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对细胞传播，并且限制了先天免疫细胞的抗病毒信号通路可以增强病毒从canineperper中从肺中释放出来。该项目的第一部分旨在研究CDV在体外对先天免疫细胞的偏振作用。在这里，CDV影响肺和血液来源巨噬细胞的表型以及单核细胞衍生的树突状细胞的成熟状态将通过流式细胞仪分析。此外，病毒介导的对巨噬细胞和树突状细胞的作用将由整个转录组分析和功能测定（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法）来表征。在第二部分中，将使用犬类空气液体界面培养物和精密的肺切片研究，将研究巨噬细胞极性和树突状细胞成熟对CDV细胞进入的影响以及调制的先天免疫细胞将CDV传输到气道上皮的能力。此外，还将确定病毒诱导的细胞质作用和超微结构变化，例如睫状病理学和凋亡诱导，以及感染培养物的再生能力。该研究将提供机械洞察力的CDV感染中肺先天免疫功能障碍及其对疾病发病机理的影响。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虫比病毒疾病中的治疗靶标，并预防向其他宿主传播病毒。