HUMAN COLONIC EPITHELIAL-CELLS, HT29/C1, TREATED WITH CRUDE BACTEROIDES-FRAGILIS ENTEROTOXIN DRAMATICALLY ALTER THEIR MORPHOLOGY
HUMAN COLONIC EPITHELIAL-CELLS, HT29/C1, TREATED WITH CRUDE BACTEROIDES-FRAGILIS ENTEROTOXIN DRAMATICALLY ALTER THEIR MORPHOLOGY
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DOI：
10.1128/iai.60.2.321-327.1992
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发表时间：
1992-02-01
影响因子：
3.1
通讯作者：
SACK, RB
中科院分区：
文献类型：
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作者：
WEIKEL, CS;GRIECO, FD;SACK, RB
脆弱类杆菌与家畜和人类的腹泻病有关。迄今为止，用于检测细菌肠毒素生物活性的常规组织培养和动物试验未能检测肠毒素B。脆弱的虽然肠易激综合征B. fragilis刺激羔羊和小牛结扎的肠袢、幼兔和结扎盲肠的成年兔的肠分泌，这些动物系统昂贵且复杂，这限制了它们用于鉴定肠促炎性B的有用性。fragilis菌株。使用克隆的人结肠上皮细胞系HT 29/C1，我们已经开发了一种体外检测方法，该方法在检测肠上皮B方面的敏感性为89%，特异性为100%。如通过羔羊结扎肠环测定所定义的脆弱菌菌株。用肠促炎性B的浓缩无菌培养物上清液处理的亚融合HT 29/C1细胞。fragilis菌株产生特异性和显著的形态学变化，包括细胞间附着的丧失、变圆、肿胀，以及在某些情况下的固缩。这些形态学变化最初在处理后1小时可见，并在至少前24小时内进展。这种组织培养试验在肠致病性B的流行病学研究中应被证明是有用的。fragilis，并可能有助于基础研究，以确定B。fragilis毒素及其作用机制。
Bacteroides fragilis has been associated with causation of diarrheal disease in livestock and humans. To date, conventional tissue culture and animal assays used to detect the biologic activity of bacterial enterotoxins have failed with enterotoxigenic B. fragilis. Although enterotoxigenic B. fragilis stimulates intestinal secretion in lamb and calf ligated intestinal loops, infant rabbits, and adult rabbits with ligated ceca, these animal systems are costly and complicated, which limits their usefulness for identification of enterotoxigenic B. fragilis strains. Using the cloned human colonic-epithelial-cell line HT29/C1, we have developed an in vitro assay that is 89% sensitive and 100% specific in detecting enterotoxigenic B. fragilis strains as defined by the lamb ligated-intestinal-loop assay. Subconfluent HT29/C1 cells treated with concentrated bacterium-free culture supernatants of enterotoxigenic B. fragilis strains develop specific and striking morphologic changes including loss of cell-to-cell attachments, rounding, swelling, and, in some cases, pyknosis. These morphologic changes are initially visible at 1 h after treatment and progress over at least the first 24 h. This tissue culture assay should prove useful in epidemiologic studies of enterotoxigenic B. fragilis and may facilitate basic studies to identify the B. fragilis toxin(s) and its mechanism of action.