Functional interaction of Escherichia coli heat-labile enterotoxin with blood group A-active glycoconjugates from differentiated HT29 cells.
Human colon adenocarcinoma cells (HT29-ATCC) and the clone HT29-5F7 were cultured under conditions that differentiate cells to a polarized intestinal phenotype. Differentiated cells showed the presence of junctional complexes and intercellular lumina bordered by microvilli. Intestinal brush border hydrolase activities (sucrase, aminopeptidase N, lactase and maltase) were detected mainly in differentiated HT29-ATCC cells compared with the differentiated clone, HT29-5F7. The presence of non-GM1 receptors of Escherichia coli heat-labile enterotoxin (LT-I) on both types of differentiated HT29 cells was indicated by the inability of cholera toxin B subunit to block LT-I binding to the cells. Binding of LT-I to cells, when GM1 was blocked by the cholera toxin B subunit, was characterized by an increased number of LT-I receptors with respect to undifferentiated control cells. Moreover, both types of differentiated cells accumulated higher amounts of cyclic AMP in response to LT-I than undifferentiated cells. Helix pomatia lectin inhibited the binding of LT-I to cells and the subsequent production of cyclic AMP. LT-I recognized blood group A-active glycosphingolipids as functional receptors in both HT29 cell lines and the active pro-sucrase form of the glycoprotein carrying A-blood group activity present in HT29-ATCC cells. These results strongly suggest that LT-I can elicit an enhanced functional response using blood group A-active glycoconjugates as additional receptors on polarized intestinal epithelial cells.
Galván
Estela M
EM
Departamento de Química Biológica-CIQUIBIC (CONICET), Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.
Roth
German A
GA
Monferran
Clara G
CG
D000017:ABO Blood-Group System; D001427:Bacterial Toxins / Q000378:metabolism*; D002454:Cell Differentiation; D000242:Cyclic AMP / Q000494:pharmacology*; D004768:Enterotoxins / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D006001:Glycoconjugates / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission
D000017:ABO Blood-Group System; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D006001:Glycoconjugates; C027538:heat-labile enterotoxin, E coli; D000242:Cyclic AMP