Oehm Andreas W, Esteves Blandina I Oliveira, Hetzel Udo, Alves Marco P, Schnyder Manuela
Institute of Parasitology, University of Zurich, Zurich, Switzerland.
Graduate School for Cellular and Biomedical Sciences, Bern, Switzerland.
Sci Rep. 2025 Mar 22;15(1):9883. doi: 10.1038/s41598-025-94033-x.
The airway epithelium represents a central barrier against pathogens and toxins while playing a crucial role in modulating the immune response within the upper respiratory tract. Understanding these mechanisms is particularly relevant for red foxes (Vulpes vulpes), which serve as reservoirs for various zoonotic pathogens like rabies or the fox tapeworm (Echinococcus multilocularis). The study aimed to develop, establish, and validate an air-liquid interface (ALI) organoid model of the fox respiratory tract using primary airway epithelial cells isolated from the tracheas and main bronchi of hunted red foxes. The resulting ALI cultures exhibited a structurally differentiated, pseudostratified epithelium, characterised by ciliated cells, mucus secretion, and tight junctions, as confirmed through histological and immunohistochemical analysis. Functional assessments using a paracellular permeability assay and measurement of transepithelial electrical resistance, demonstrated a tight epithelial barrier. The potential of model's utility for studying innate immune responses to respiratory infections was validated by exposing the cultures to lipopolysaccharide, phorbol-12-myristate-13-acetate and ionomycin, and nematode somatic antigens. Quantitative PCR revealed notable changes in the expression of pro-inflammatory cytokines TNF and IL-33. This in vitro model represents a significant advancement in respiratory research for non-classical species that may act as important wildlife reservoirs for a range of zoonotic pathogens.
气道上皮是抵御病原体和毒素的主要屏障，同时在调节上呼吸道的免疫反应中发挥关键作用。了解这些机制对于赤狐（Vulpes vulpes）尤为重要，因为它们是狂犬病或狐绦虫（多房棘球绦虫）等多种人畜共患病原体的宿主。该研究旨在利用从被捕猎的赤狐气管和主支气管分离的原代气道上皮细胞，开发、建立并验证狐呼吸道的气液界面（ALI）类器官模型。通过组织学和免疫组织化学分析证实，所得的ALI培养物呈现出结构分化的假复层上皮，其特征为纤毛细胞、黏液分泌和紧密连接。使用跨细胞通透性测定和跨上皮电阻测量进行的功能评估表明存在紧密的上皮屏障。通过将培养物暴露于脂多糖、佛波醇-12-肉豆蔻酸酯-13-乙酸酯和离子霉素以及线虫体抗原，验证了该模型用于研究对呼吸道感染的固有免疫反应的效用潜力。定量PCR显示促炎细胞因子TNF和IL-33的表达有显著变化。这种体外模型代表了非经典物种呼吸道研究的重大进展，这些物种可能是一系列人畜共患病原体的重要野生动物宿主。
