Coinfection Mechanisms ofCampylobacter andEscherichia coli in Human and Chicken Epithelial cells
Coinfection Mechanisms ofCampylobacter andEscherichia coli in Human and Chicken Epithelial cells
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人和鸡上皮细胞中弯曲杆菌和大肠杆菌的共感染机制
DOI：
10.1099/acmi.fis2019.po0165
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发表时间：
2020
影响因子：
--
通讯作者：
Hanford T
中科院分区：
文献类型：
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作者：
Hanford T
空肠弯曲菌是世界上最常见的食源性致病菌。弯曲杆菌的致病机制涉及人和鸡的肠上皮细胞屏障的易位，以前的工作表明，E。大肠杆菌（ExPEC/APEC）可能促进这一过程。本研究的目的是确定这种关系可能对两种微生物的入侵和粘附潜力的影响，使用我们最近发表的人类和鸟类的体外模型。C. jejuni和E.使用源自胃肠道的人（Caco-2）和禽（8 E-11）上皮细胞系测量大肠杆菌菌株，并通过落射荧光和共聚焦显微镜进行表征。进行粘附和侵袭测定以确定不同细菌的致病性。Caco-2和8E-11连同细菌菌株的代谢活性，使用亚甲蓝测定。共聚焦和落射荧光显微镜确定Caco-2细胞中细胞角蛋白的强存在，而在8E-11细胞中检测到弱至中等信号。庆大霉素在所有菌株中的最佳剂量和时间为0.02mg/ml，90分钟，不影响上皮细胞的代谢活性。当暴露于人类和禽类细胞类型时，发现细菌的粘附/侵入潜力存在显著差异。C. jejuni（11168）和E.大肠杆菌（K12）中，K12的存在对11168的活性具有负面影响。这里描述的模型将提供机会，以提高我们的弯曲杆菌在人类和鸡的入侵机制的理解，使改进的策略，以否定这些后果可能会被设计。
Campylobacter jejuniis the world’s most common food-borne pathogen. Campylobacterpathogenesis involves translocation across the intestinal epithelial cell barrier in both humans and chickens and previous work has suggested that strains of E. coli (ExPEC / APEC) may facilitate this process. This study aims to determine the effect that this relationship may have upon the invasive and adhesive potential of the two microbes using our recently published human and avian in vitromodel. Invasiveness of C. jejuni and E. coli strains were measured using human (Caco-2) and avian (8E-11) epithelial cell lines derived from the gastrointestinal tract and were characterised by epifluorescent and confocal microscopy. Adhesion and invasion assays were carried out to determine the pathogenicity of the different bacteria. Metabolic activity of Caco-2 and 8E-11 together with bacterial strains using alamar blue assay. Confocal and epifluorescence microscopy determined the strong presence of cytokeratin in Caco-2 cells whilst weak to medium signals were detected in 8E-11 cells. Optimal doses and times for of gentamicin across all strains was 0.02mg/ml for 90 minutes which did not affect metabolic activity of epithelial cells. Significant diversity was found in the adhesive/invasive potential of bacteria when exposed to human and avian cell types. The metabolic rate of C. jejuni (11168) and E. coli (K12) was investigated with the presence of K12 having negative impacts upon the activity of 11168. The model described here will provide opportunity to improve our understanding of Campylobacterinvasion mechanisms in human and chickens so that improved strategies to negate these consequences may be designed.