Sparber Florian, LeibundGut-Landmann Salomé
Section of Immunology, Vetsuisse Faculty, University of Zürich;
J Vis Exp. 2019 Nov 6(153). doi: 10.3791/60175.
Animal models are crucial for infectious disease research. They provide an important basis for analyzing the full spectrum of interactions that occur between microbes and their host in vivo in a tissue-specific manner. Pathogenic fungi are increasingly recognized as a serious threat for humans and exploiting such infection models have greatly improved our understanding of fungal pathogenicity. Species of the genus Malassezia are the most abundant fungi of the human skin microbiota and they are also associated with the development of severe inflammatory skin disorders such as seborrheic dermatitis and atopic dermatitis. However, a causative link between Malassezia and disease pathogenesis remains unknown, a fact that can be attributed to the poor knowledge of the complex crosstalk of Malassezia with the skin immune system. This protocol describes the establishment of an experimental mouse model that allows studying the interaction of Malassezia with the mammalian skin in vivo. It outlines the method for cultivating Malassezia spp. under laboratory conditions, how to infect the murine skin with Malassezia spp. and how to assess the outcome of infection by means of the skin inflammation and fungal burden analyses. The model described here works in fully immunocompetent animals and does not rely on immune suppressive or antibiotic pretreatment of the animals. It is furthermore adaptable to virtually all genetically modified mouse strains and can be combined with other skin disease models. These features make this infection model a very powerful tool for studying in detail the innate and adaptive immune response of the host against Malassezia in the skin in vivo.
动物模型对于传染病研究至关重要。它们为以组织特异性方式分析微生物与其宿主体内发生的全谱相互作用提供了重要依据。致病性真菌日益被视为对人类的严重威胁,利用此类感染模型极大地增进了我们对真菌致病性的理解。马拉色菌属物种是人类皮肤微生物群中最丰富的真菌,它们还与严重炎症性皮肤病如脂溢性皮炎和特应性皮炎的发生有关。然而,马拉色菌与疾病发病机制之间的因果联系仍不明确,这一事实可归因于对马拉色菌与皮肤免疫系统复杂相互作用的了解不足。本方案描述了一种实验性小鼠模型的建立,该模型可用于研究马拉色菌与哺乳动物皮肤在体内的相互作用。它概述了在实验室条件下培养马拉色菌属物种的方法、如何用马拉色菌属物种感染小鼠皮肤以及如何通过皮肤炎症和真菌负荷分析评估感染结果。这里描述的模型适用于完全免疫 competent 的动物,不依赖于对动物进行免疫抑制或抗生素预处理。此外,它几乎适用于所有转基因小鼠品系,并且可以与其他皮肤病模型相结合。这些特性使这种感染模型成为详细研究宿主对皮肤内马拉色菌的固有和适应性免疫反应的非常强大的工具。