引用

艰难梭菌是一种重要的医院病原体，它会产生毒素，导致危及生命的腹泻和结肠炎。毒素结合上皮受体，促进肌动蛋白细胞骨架的崩溃。艰难梭菌毒素活性通常在癌症来源和永生细胞系中进行研究。然而，这些模型的生物学相关性是有限的。此外，尚无模型可用于检查艰难梭菌诱导的肠炎，这是一个研究不足的健康问题。我们假设人肠类肠(HIEs)表达毒素受体，并提供了一个新的模型来解剖小肠中的艰难梭菌细胞毒性。我们生成了活组织切片来源的空肠HIE和Vero细胞，它们稳定表达LifeAct-Ruby, F-actin的荧光标记，通过活细胞显微镜监测actin细胞骨架重排。成像分析显示，来自致病性艰难梭菌菌株的毒素以菌株依赖的方式引起细胞转圆，HIEs对毒素a (TcdA)比毒素B (TcdB)敏感10倍。通过定量PCR，我们矛盾地发现，与传统使用的细胞系相比，HIEs表达了更多的毒素受体mRNA，但对毒素的敏感性却降低了。我们推断，这些差异可能是由HIEs培养中存在的黏蛋白等成分解释的，而这些成分在永生化细胞系中是不存在的。 Addition of human-derived mucin 2 (MUC2) to Vero cells delayed cell rounding, indicating that mucus serves as a barrier to toxin-receptor binding. This work highlights that investigation of C. difficile infection in that HIEs can provide important insights into the intricate interactions between toxins and the human intestinal epithelium.NEW & NOTEWORTHY In this article, we developed a novel model of Clostridioides difficile-induced enteritis using jejunal-derived human intestinal enteroids (HIEs) transduced with fluorescently tagged F-actin. Using live-imaging, we identified that jejunal HIEs express high levels of TcdA and CDT receptors, are more sensitive to TcdA than TcdB, and secrete mucus, which delays toxin-epithelial interactions. This work also optimizes optically clear C. difficile-conditioned media suitable for live-cell imaging.