Belley A, Keller K, Grove J, Chadee K
Institute of Parasitology, McGill University, Quebec, Canada.
Gastroenterology. 1996 Dec;111(6):1484-92. doi: 10.1016/s0016-5085(96)70009-4.
BACKGROUND & AIMS: Colonic mucins secreted by goblet cells protect the colon by preventing the attachment of enteric pathogens to the epithelium. Entamoeba histolytica overcomes this protective barrier and causes ulcerations, allowing the parasite to disseminate to the liver and form abscesses. An in vitro model is used to study the interaction between E. histolytica and colonic mucins.
Secretory mucins from the colonic adenocarcinoma cell line LS174T were collected and their functions assessed by their ability to inhibit amebic adherence to target cells and killing. The cytoprotective effect of mucus against E. histolytica cytolysis of LS174T monolayers was studied at 37 degrees C.
Sepharose 4B column chromatography, metabolic labeling with [3H]glucosamine, cesium chloride density gradient centrifugation, and amino acid and carbohydrate compositional analysis revealed that LS174T cell mucins were typical of native colonic mucins. Mucin O-linked oligosaccharides bound to and inhibited the adherence of amebae to Chinese hamster ovary cells. E. histolytica killing of Chinese hamster ovary cell monolayers occurred rapidly, whereas killing of LS174T monolayers with an intact mucus layer was significantly retarded.
Our results show that colonic mucins serve as the first line of host defense against amebic invasion and provide a useful model to study pathogen-mucin interactions.
杯状细胞分泌的结肠黏蛋白通过防止肠道病原体附着于上皮细胞来保护结肠。溶组织内阿米巴克服了这一保护屏障并导致溃疡形成,使该寄生虫得以扩散至肝脏并形成脓肿。体外模型用于研究溶组织内阿米巴与结肠黏蛋白之间的相互作用。
收集结肠腺癌细胞系LS174T分泌的黏蛋白,并通过其抑制阿米巴对靶细胞的黏附和杀伤的能力来评估其功能。在37℃下研究黏液对溶组织内阿米巴对LS174T单层细胞的细胞溶解作用的细胞保护作用。
琼脂糖4B柱层析、用[3H]葡糖胺进行代谢标记、氯化铯密度梯度离心以及氨基酸和碳水化合物组成分析显示,LS174T细胞黏蛋白是天然结肠黏蛋白的典型代表。黏蛋白O-连接寡糖结合并抑制阿米巴对中国仓鼠卵巢细胞的黏附。溶组织内阿米巴对中国仓鼠卵巢细胞单层的杀伤迅速发生,而对具有完整黏液层的LS174T单层细胞的杀伤则明显延迟。
我们的结果表明,结肠黏蛋白是宿主抵御阿米巴入侵的第一道防线,并为研究病原体与黏蛋白的相互作用提供了一个有用的模型。