Schaller Martin, Zakikhany Katherina, Naglik Julian R, Weindl Günther, Hube Bernhard
Department of Dermatology, Eberhard Karls University, Liebermeisterstr, 25, D-72076 Tuebingen, Germany.
Nat Protoc. 2006;1(6):2767-73. doi: 10.1038/nprot.2006.474.
This protocol describes the setup, maintenance and characteristics of models of epithelial Candida infections based on well-established three-dimensional organotypic tissues of human oral and vaginal mucosa. Infection experiments are highly reproducible and can be used for the direct analysis of pathogen-epithelial cell interactions. This allows detailed investigations of Candida albicans wild type or mutant strain interaction with epithelial tissue or the evaluation of the host immune response using histological, biochemical and molecular methods. As such, the models can be utilized as a tool to investigate cellular interactions or protein and gene expression that are not complicated by non-epithelial factors. To study the impact of innate immunity or the antifungal activity of natural and non-natural compounds, the mucosal infection models can be supplemented with immune cells, antimicrobial agents or probiotic bacteria. The model requires at least 3 days to be established and can be maintained thereafter for 2-4 days.
本方案描述了基于成熟的人口腔和阴道黏膜三维器官型组织的上皮念珠菌感染模型的建立、维护及特征。感染实验具有高度可重复性,可用于直接分析病原体与上皮细胞的相互作用。这使得能够使用组织学、生物化学和分子方法详细研究白色念珠菌野生型或突变株与上皮组织的相互作用,或评估宿主免疫反应。因此,这些模型可作为一种工具,用于研究不受非上皮因素干扰的细胞相互作用或蛋白质和基因表达。为了研究先天免疫的影响或天然及非天然化合物的抗真菌活性,黏膜感染模型可补充免疫细胞、抗菌剂或益生菌。该模型至少需要3天来建立,此后可维持2 - 4天。