Basheer Faiza, Liongue Clifford, Ward Alister C
School of Medicine, Deakin University, Geelong, Victoria, Australia.
Centre for Molecular and Medical Research, Deakin University, Geelong, Victoria, Australia.
Bio Protoc. 2020 Mar 5;10(5):e3536. doi: 10.21769/BioProtoc.3536.
The study of host-pathogen interactions has improved our understanding of both pathogenesis and the response of the host to infection, including both innate and adaptive responses. Neutrophils and macrophages represent the first line of innate host defense against any infection. The zebrafish is an ideal model to study the response of these cells to a variety of pathogens. Zebrafish possess both neutrophils and macrophages exhibiting similar defense mechanisms to their human counterparts. The transparency of zebrafish embryos greatly facilitates tracking of infection dynamics in a non-invasive manner at high-resolution using labelled pathogens, while immune cells can also be labelled transgenically to enable even more in-depth analysis. Here we describe a procedure for performing a bacterial infection assay in zebrafish embryos using fluorescently-labelled bacteria and demonstrate the monitoring and quantification of the infection kinetics. Of note, this procedure helps in understanding the functional role of genes that are important in driving the innate immune response.
宿主-病原体相互作用的研究增进了我们对发病机制以及宿主对感染反应的理解,包括先天性和适应性反应。中性粒细胞和巨噬细胞是宿主抵御任何感染的先天性防御的第一道防线。斑马鱼是研究这些细胞对多种病原体反应的理想模型。斑马鱼拥有中性粒细胞和巨噬细胞,其防御机制与人类对应细胞相似。斑马鱼胚胎的透明性极大地便于使用标记病原体以高分辨率、非侵入性方式追踪感染动态,同时免疫细胞也可以通过转基因进行标记,从而实现更深入的分析。在此,我们描述了一种在斑马鱼胚胎中使用荧光标记细菌进行细菌感染检测的方法,并展示了对感染动力学的监测和定量。值得注意的是,该方法有助于理解在驱动先天性免疫反应中起重要作用的基因的功能。