这项研究解决了用于研究肠出血性大肠杆菌 (EHEC) 感染的可转化体外模型的空白，特别是与犬类和人类健康相关的感染。众所周知，肠出血性大肠杆菌会诱发狗的急性结肠炎，导致出血性腹泻和溶血性尿毒症综合征等症状，与人类中观察到的症状类似。然而，由于缺乏在这两个物种中复制肠出血性大肠杆菌引起的临床疾病的有效模型，了解病理生理学和制定治疗策略一直具有挑战性。我们的方法涉及使用来自客户拥有的健康狗的肠道组织开发源自类结肠的单层细胞。这些单层细胞暴露于肠出血性大肠杆菌，并通过多种技术评估肠出血性大肠杆菌的影响，包括跨上皮电阻 (TEER) 测量、连接蛋白和粘液的免疫荧光染色以及用于形态学分析的扫描电子显微镜。用盐水改良培养物，旨在防止细菌过度生长，保持屏障完整性和细胞分化。 EHEC 感染导致 TEER 和 ZO-1 表达显着降低，但不会导致 E-钙粘蛋白水平或粘液产生显着降低。此外，肠出血性大肠杆菌引起肿瘤坏死因子-α 的产生显着增加，突显了与非致病性大肠杆菌相比，其对犬肠上皮细胞的独特影响。这些发现与患有肠出血性大肠杆菌肠病的狗和人类的体内观察结果非常相似，验证了犬类结肠源性单层系统作为研究肠出血性大肠杆菌和其他潜在临床重要肠道病原体中宿主-病原体相互作用的转化模型。这项研究开发了一种新模型更好地了解肠出血性大肠杆菌 (EHEC) 感染，这是一种影响狗和人类的严重细菌性疾病，其特征是出血性腹泻和溶血性尿毒症综合征等症状。传统的研究模型无法模拟这种疾病在患者中的表现。我们的研究使用健康狗的肠道组织来创建细胞层，称为结肠源性单层细胞，然后将其暴露于肠出血性大肠杆菌中。我们使用多种技术评估了细菌造成的损害，观察到与实际疾病病例类似的显着变化。该模型被证明可以有效地复制宿主和病原体之间的相互作用，标志着了解肠出血性大肠杆菌的影响和开发治疗方法的重要一步。这种源自犬类集落的单层系统不仅弥补了当前研究中的一个关键空白，而且还为研究影响犬类和人类健康的其他肠道病原体提供了一个有前途的平台。

This study addresses the gap in translatable in vitro models for investigating Enterohemorrhagic E. coli (EHEC) infections, particularly relevant to both canine and human health. EHEC is known to induce acute colitis in dogs, leading to symptoms like hemorrhagic diarrhea and hemolytic uremic syndrome, similar to those observed in humans. However, understanding the pathophysiology and developing treatment strategies have been challenging due to the lack of effective models that replicate the clinical disease caused by EHEC in both species. Our approach involved the development of colonoid-derived monolayers using intestinal tissues from healthy, client-owned dogs. These monolayers were exposed to EHEC, and the impact of EHEC was assessed through several techniques, including trans-epithelial electrical resistance (TEER) measurement, immunofluorescence staining for junction proteins and mucus, and scanning electron microscopy for morphological analysis. Modified culture with saline, which was intended to prevent bacterial overgrowth, maintained barrier integrity and cell differentiation. EHEC infection led to significant decreases in TEER and ZO-1 expression, but not in E-cadherin levels or mucus production. In addition, EHEC elicited a notable increase in tumor necrosis factor-alpha production, highlighting its distinct impact on canine intestinal epithelial cells compared to non-pathogenic E. coli. These findings closely replicate in vivo observations in dogs and humans with EHEC enteropathy, validating the canine colonoid-derived monolayer system as a translational model to study host-pathogen interactions in EHEC and potentially other clinically significant enteric pathogens.This study develops a new model to better understand Enterohemorrhagic E. coli (EHEC) infections, a serious bacterial disease affecting both dogs and humans, characterized by symptoms such as hemorrhagic diarrhea and hemolytic uremic syndrome. Traditional research models have fallen short of mimicking how this disease manifests in patients. Our research used intestinal tissues from healthy dogs to create layers of cells, known as colonoid-derived monolayers, which we then exposed to EHEC. We assessed the damage caused by the bacteria using several techniques, observing significant changes similar to those seen in actual cases of the disease. The model proved effective in replicating the interaction between the host and the pathogen, marking an important step toward understanding EHEC's effects and developing treatments. This canine colonoid-derived monolayer system not only bridges a crucial gap in current research but also offers a promising platform for studying other enteric pathogens affecting both canine and human health.