Hochstrasser Ramon, Hilbi Hubert
Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland.
Methods Mol Biol. 2019;1921:79-89. doi: 10.1007/978-1-4939-9048-1_5.
The amoeba-resistant bacterium Legionella pneumophila infects humans through aerosols and thereby can cause a life-threatening pneumonia termed Legionnaires' disease. In the environment L. pneumophila forms and colonizes biofilms, which usually comprise complex multispecies communities. In these biofilms L. pneumophila persists and replicates intracellularly in protozoa, such as the amoeba Acanthamoeba castellanii. The interactions between sessile L. pneumophila in biofilms and their natural protozoan hosts are not understood on a molecular level. Here, we describe a method to visualize by confocal microscopy the formation and architecture of mono-species L. pneumophila biofilms. Furthermore, we describe and quantify the migration or "grazing" of A. castellanii in the biofilm. This allows investigating on a molecular and cellular level L. pneumophila biofilm formation and Legionella-amoeba interactions within biofilms.
抗变形虫的嗜肺军团菌通过气溶胶感染人类，从而可能引发一种危及生命的肺炎，称为军团病。在环境中，嗜肺军团菌形成并定殖于生物膜，生物膜通常由复杂的多物种群落组成。在这些生物膜中，嗜肺军团菌在原生动物（如卡氏棘阿米巴变形虫）内持续存在并在细胞内复制。生物膜中固着的嗜肺军团菌与其天然原生动物宿主之间的相互作用在分子水平上尚不清楚。在此，我们描述了一种通过共聚焦显微镜观察单物种嗜肺军团菌生物膜形成和结构的方法。此外，我们描述并量化了卡氏棘阿米巴变形虫在生物膜中的迁移或“捕食”行为。这使得能够在分子和细胞水平上研究嗜肺军团菌生物膜的形成以及生物膜内军团菌与变形虫之间的相互作用。
