Bernet-Camard M F, Duigou F, Kernéis S, Coconnier M H, Servin A L
CJF 94.07 INSERM, UFR de Pharmacie Paris XI, Châtenay-Malabry, France.
Infect Immun. 1997 Apr;65(4):1299-306. doi: 10.1128/iai.65.4.1299-1306.1997.
The association of enterotoxigenic Escherichia coli expressing colonization factor antigen I (CFA/I) with the cultured human colon adenocarcinoma cell, a model of the mature enterocyte of the small intestine, is dependent on the binding of CFA/I to a brush border-associated component. Binding of the purified radiolabeled [125I]CFA/I- and 14C-labeled CFA/I-positive bacteria could be displaced by an increasing concentration of unlabeled CFA/I. Moreover, we showed that expression of the specific CFA/I binding developed as a function of cell differentiation in Caco-2 cells, whereas expression of the nonspecific binding did not. Expression of the brush border differentiation-associated component acting as a binding site for CFA/I was up-regulated by glucose. Indeed, the enterocyte-like HT-29 glc- cell subpopulation not expressing the CFA/I binding site when cultured in dialyzed serum and hexose-free medium regained the ability to bind CFA/I when the cells were returned to culture medium containing glucose. Furthermore, expression of the brush border-associated CFA/I binding site in the enterocyte-like Caco-2 cells was repressed when the cells were cultured in hexose-free conditions.
表达定植因子抗原I(CFA/I)的产肠毒素大肠杆菌与培养的人结肠腺癌细胞(一种小肠成熟肠上皮细胞的模型)的结合,取决于CFA/I与刷状缘相关成分的结合。纯化的放射性标记的[125I]CFA/I和14C标记的CFA/I阳性细菌的结合可被浓度不断增加的未标记CFA/I所取代。此外,我们发现,在Caco-2细胞中,特异性CFA/I结合的表达是细胞分化的函数,而非特异性结合的表达则不是。作为CFA/I结合位点的刷状缘分化相关成分的表达被葡萄糖上调。实际上,当在透析血清和无己糖培养基中培养时不表达CFA/I结合位点的肠上皮样HT-29 glc-细胞亚群,当细胞重新培养于含葡萄糖的培养基中时,恢复了结合CFA/I的能力。此外,当肠上皮样Caco-2细胞在无己糖条件下培养时,刷状缘相关的CFA/I结合位点的表达受到抑制。