Izhar M, Nuchamowitz Y, Mirelman D
Infect Immun. 1982 Mar;35(3):1110-8. doi: 10.1128/iai.35.3.1110-1118.1982.
Guinea pig colonic epithelial cells released by treating sections of the colon with solutions containing EDTA, dithiothreitol, and citrate avidly adhered Shigella flexneri bacteria. Separation of the intestinal cells from nonbound bacteria was achieved by differential sedimentation on a Percoll gradient. Adherence of S. flexneri to the colonic cells was Ca2+ (1 mM) and time dependent. The pH optimum was pH 6.2, and almost no attachment (less than 5%) was observed at low temperature (4 degrees C). The average number of bacteria which bound to colonic cells was 70 bacteria per cell, whereas attachment to cells isolated from the ileum region was 6 bacteria per cell. Colonic cells obtained from the intestine of rabbits or rats did not adhere Shigella. Adherence to guinea pig colonic cells was inhibited (50%) by several carbohydrates, such as 0.1% fucose or 0.5% glucose, as well as by a lipopolysaccharide preparation (10 micrograms /ml) isolated from S. flexneri. Fixation of the bacteria with glutaraldehyde or preincubation of the bacteria with lectins or proteolytic enzymes did not affect their adherence. Proteolytic digestions or fixation of the epithelial cells, as well as pretreatments with lipopolysaccharide or fucose solutions, abolished their ability to adhere bacteria. These results indicate that a carbohydrate-binding substance on the surface of guinea pig colonic epithelial cells is responsible for the attachment of the Shigella bacilli.
用含有乙二胺四乙酸（EDTA）、二硫苏糖醇和柠檬酸盐的溶液处理结肠切片后释放出的豚鼠结肠上皮细胞，能强烈黏附福氏志贺氏菌。通过在Percoll梯度上进行差速沉降，可将肠道细胞与未结合的细菌分离。福氏志贺氏菌对结肠细胞的黏附依赖于Ca2+（1 mM）且具有时间依赖性。最适pH为6.2，在低温（4℃）下几乎没有黏附（少于5%）现象。黏附到结肠细胞上的细菌平均数量为每个细胞70个细菌，而黏附到从回肠区域分离出的细胞上的细菌数量为每个细胞6个。从兔或大鼠肠道获得的结肠细胞不黏附志贺氏菌。几种碳水化合物，如0.1%岩藻糖或0.5%葡萄糖，以及从福氏志贺氏菌中分离出的脂多糖制剂（10微克/毫升），可抑制（50%）对豚鼠结肠细胞的黏附。用戊二醛固定细菌，或用凝集素或蛋白水解酶对细菌进行预孵育，均不影响其黏附。对上皮细胞进行蛋白水解消化或固定，以及用脂多糖或岩藻糖溶液进行预处理，均会消除其黏附细菌的能力。这些结果表明，豚鼠结肠上皮细胞表面的一种碳水化合物结合物质负责志贺氏杆菌的黏附。
