Hachfi Salma, Benguettat Olivia, Gallet Armel
Universite Cote d'Azur, CNRS, INRA, ISA, France.
Institut Sophia Agrobiotech, UMR CNRS 7254/INRA 1355/UNS, 400 route des Chappes, BP 167, 06903 Sophia Antipolis Cedex, France.
Bio Protoc. 2019 May 20;9(10):e3246. doi: 10.21769/BioProtoc.3246.
The intestine is endowed with an innate immune system that is required to fight any exogenous bacteria that are swallowed along with the food. The first line of defense that is mounted by the gut epithelium is the release of immune Reactive Oxygen Species (ROS), such as hypochlorous acid (HOCl), into the lumen. HOCl is produced within 1.5 h of bacterial ingestion and is very labile once released. Therefore, to monitor HOCl production upon ingestion of allochthonous bacteria, one needs a detection system that can quickly and efficiently detect HOCl production in the intestine. While most of the ROS-sensitive probes available in the market detect all kinds of ROS without any distinction, the R19-S fluorescent probe has been developed to specifically detect HOCl. Here, we describe a protocol to monitor HOCl production using this probe in the gut lumen of adult upon ingestion of the opportunistic bacteria .
肠道具有先天性免疫系统，该系统对于抵抗随食物一同摄入的任何外源细菌至关重要。肠道上皮细胞启动的第一道防线是将免疫活性氧（ROS），如次氯酸（HOCl）释放到肠腔中。HOCl在摄入细菌后1.5小时内产生，一旦释放便极不稳定。因此，为了监测摄入外源细菌后HOCl的产生情况，需要一种能够快速、高效地检测肠道中HOCl产生的检测系统。虽然市场上大多数ROS敏感探针不加区分地检测各种ROS，但R19-S荧光探针已被开发用于特异性检测HOCl。在此，我们描述了一种使用该探针监测成年个体摄入机会性细菌后肠道腔中HOCl产生情况的方案。
