Salmonella-infected crypt-derived intestinal organoid culture system for host-bacterial interactions.
Salmonella-infected crypt-derived intestinal organoid culture system for host-bacterial interactions.
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DOI：
10.14814/phy2.12147
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发表时间：
2014-09-01
影响因子：
2.5
通讯作者：
Sun J
中科院分区：
文献类型：
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作者：
Zhang YG;Wu S;Xia Y;Sun J
由于缺乏合适的肠上皮细胞培养系统，肠内细菌-上皮细胞相互作用的体外分析受到阻碍。在这里，我们报告了一个新的实验模型，使用类器官培养系统研究沙门氏菌感染后细菌-上皮相互作用的病理生理学。使用隐窝衍生的小鼠肠道类器官，我们能够可视化沙门氏菌的侵袭性和类器官的形态学变化。重要的是，我们报告了细菌诱导的受感染类器官上皮紧密连接的破坏。此外，我们通过类器官中NF-κB通路的激活显示了炎症反应。此外，我们的蛋白质印迹、PCR和免疫荧光数据表明，沙门氏菌感染显著降低了干细胞标志物（Lgr 5和Bmi 1）（使用GFP标记的Lgr 5类器官测定）。这是第一次，我们创建了一个模型系统，概括了沙门氏菌感染肠道的体内研究中的许多观察结果，包括细菌入侵，改变紧密连接，炎症反应和干细胞减少。我们已经证明，沙门氏菌感染的类器官培养系统是一种适合研究宿主-细菌相互作用的新实验模型。使用干细胞衍生的肠类器官，我们能够可视化沙门氏菌的侵袭性，类器官的形态学变化，并通过激活类器官中的NF-κB通路显示炎症反应。此外，我们发现，干细胞标志物（Lgr 5和Bmi 1）减少沙门氏菌感染。总之，我们重新创建的模型系统首次概括了沙门氏菌感染肠道体内研究的许多观察结果。因此，沙门氏菌感染的类器官培养系统是一种适合研究宿主-细菌相互作用的新实验模型。
The in vitro analysis of bacterial–epithelial interactions in the intestine has been hampered by a lack of suitable intestinal epithelium culture systems. Here, we report a new experimental model using an organoid culture system to study pathophysiology of bacterial–epithelial interactions post Salmonella infection. Using crypt‐derived mouse intestinal organoids, we were able to visualize the invasiveness of Salmonella and the morphologic changes of the organoids. Importantly, we reported bacteria‐induced disruption of epithelial tight junctions in the infected organoids. In addition, we showed the inflammatory responses through activation of the NF‐κB pathway in the organoids. Moreover, our western blot, PCR, and immunofluorescence data demonstrated that stem cell markers (Lgr5 and Bmi1) were significantly decreased by Salmonella infection (determined using GFP‐labeled Lgr5 organoids). For the first time, we created a model system that recapitulated a number of observations from in vivo studies of the Salmonella‐infected intestine, including bacterial invasion, altered tight junctions, inflammatory responses, and decreased stem cells. We have demonstrated that the Salmonella‐infected organoid culture system is a new experimental model suitable for studying host–bacterial interactions. Using stem cell‐derived intestinal organoids, we were able to visualize the invasiveness of Salmonella, morphologic changes of the organoids, and showed the inflammatory responses through activating the NF‐κB pathway in the organoids. Moreover, we show that stem cell markers (Lgr5 and Bmi1) were decreased by Salmonella infection. Taken together, the model system we have recreated recapitulates for the first time a number of observations from in vivo studies of Salmonella‐infected gut. Thus, Salmonella‐infected organoid culture system is a new experimental model suitable for studying host–bacterial interactions.