Shteindel Nimrod, Gutman Danielle, Atzmon Gil, Gerchman Yoram
Department of Evolutionary and Environmental Biology, Haifa University, Tivon, Israel.
Department of Human Biology, University of Haifa, Haifa 3498838, Israel.
Biol Methods Protoc. 2023 Jul 26;8(1):bpad014. doi: 10.1093/biomethods/bpad014. eCollection 2023.
Bacterial adhesion to tissue is the starting point for many pathogenic processes and beneficial interactions. The dynamics and speed of adhesion (minutes) make high-resolution temporal kinetic data important, but this capability is absent from the current toolset. We present a high-throughput method with a second-to-minute kinetic resolution, testing the adhesion of PAO1 wild-type, flagella-, pili-, and quorum-sensing mutants to human embryonic kidney (HEK293) cells. Adhesion rates were in good correlation with HEK293 confluence, and the ways in which various bacterial mutations modified adhesion patterns are in agreement with the published literature. This simple assay can facilitate drug screening and treatment development as well as provide a better understanding of the interactions of pathogenic and probiotic bacteria with tissues, allowing the design of interventions and prevention treatments.
细菌对组织的黏附是许多致病过程和有益相互作用的起点。黏附的动力学和速度(以分钟计)使得高分辨率的时间动力学数据变得重要,但当前的工具集缺乏这种能力。我们提出了一种具有每分钟动力学分辨率的高通量方法,用于测试PAO1野生型、鞭毛突变体、菌毛突变体和群体感应突变体对人胚肾(HEK293)细胞的黏附。黏附率与HEK293细胞汇合度具有良好的相关性,并且各种细菌突变改变黏附模式的方式与已发表的文献一致。这种简单的检测方法可以促进药物筛选和治疗开发,同时更好地理解致病细菌和益生菌与组织的相互作用,从而设计干预措施和预防治疗方法。