Generation of self-replicating airway organoids from the cave nectar bat <i>Eonycteris spelaea</i> as a model system for studying host–pathogen interactions in the bat airway epithelium

Bats are reservoir hosts for various zoonotic viruses with pandemdic potential in humans and livestock. <i>In vitro</i> systems for studying bat host–pathogen interactions are of significant interest. Here, we establish protocols to generate bat airway organoids (AOs) and airway epithelial cells differentiated at the air–liquid interface (ALI-AECs) from tracheal tissues of the cave-nectar bat <i>Eonycteris spelaea</i>. In particular, we describe steps which enable laboratories that do not have access to live bats to perform extended experimental work upon procuring an initial batch of bat primary airway tissue. Complete mucociliary differentiation required treatment with IL-13. <i>E. spelaea</i> ALI-AECs supported productive infection with PRV3M, an orthoreovirus for which Pteropodid bats are considered the reservoir species. However, these ALI-AECs did not support SARS-CoV-2 infection, despite <i>E. spelaea</i> ACE2 receptor being capable of mediating SARS-CoV-2 spike pseudovirus entry. This work provides critical model systems for assessing bat species-specific virus susceptibility and the reservoir likelihood for emerging infectious agents.

蝙蝠是多种具备大流行潜力的人畜共患病毒的自然储存宿主，此类病毒可感染人类与家畜。用于研究蝙蝠宿主-病原体互作的体外（in vitro）模型具有重要研究价值。本研究建立了从穴居食花蜜短鼻犬蝠（*Eonycteris spelaea*）的气管组织中制备蝙蝠气道类器官（airway organoids, AOs）以及气液界面分化气道上皮细胞（air-liquid interface differentiated airway epithelial cells, ALI-AECs）的实验方案。尤为关键的是，本研究详细描述了无需直接接触活体蝙蝠即可开展扩展实验的操作流程：仅需获取首批蝙蝠原代气道组织即可完成后续实验。完全的黏膜纤毛分化需经白细胞介素-13（IL-13）处理。短鼻犬蝠ALI-AECs可支持PRV3M的增殖性感染，该病毒属于正呼肠孤病毒（orthoreovirus），狐蝠科蝙蝠被认定为其自然储存宿主。然而，尽管短鼻犬蝠的血管紧张素转换酶2（angiotensin-converting enzyme 2, ACE2）受体能够介导新型冠状病毒（SARS-CoV-2）刺突假病毒的入侵，但此类ALI-AECs并不能支持SARS-CoV-2的感染。本研究构建的模型系统可为评估蝙蝠物种特异性病毒易感性以及新兴病原体的储存宿主潜力提供关键实验工具。

搜集汇总

背景与挑战

背景概述

该数据集描述了从洞穴果蝠（Eonycteris spelaea）生成自复制气道类器官和气液界面分化上皮细胞的协议，旨在建立研究蝙蝠气道上皮宿主-病原体相互作用的体外模型系统。实验表明，这些模型支持PRV3M病毒感染，但不支持SARS-CoV-2感染，为评估蝙蝠物种特异性病毒易感性和新兴病原体储存潜力提供了关键工具。数据集涵盖了相关资助、类别和关键词信息，适用于医学、生态学和病毒学等领域的研究。