title: Clostridium difficile toxin A binding to human intestinal epithelial cells.
abstract: Clostridium difficile radiolabelled toxin A ([3H]-toxin A) bound to human duodenal and colonic epithelial cells isolated from endoscopic biopsies. Binding was greater at 4 degrees C than 37 degrees C, consistent with the thermal binding characteristic of toxin A to a carbohydrate moiety. At 37 degrees C colonic cells bound significantly more [3H]-toxin A than duodenal cells. The amount of [3H]-toxin A binding varied considerably between individuals. [3H]-toxin A was displaced by unlabelled toxin A by 50% for duodenal cells and 70% for colonic cells with 94.3 nM unlabelled toxin A. Low non-displacable binding was observed in some samples at 4 degrees C and 37 degrees C, suggesting that these cells came from individuals incapable of specifically binding toxin. Pre-treating cells with alpha- or beta-galactosidases to cleave terminal alpha- and beta-galactose residues reduced [3H]-toxin A binding. There was also a reduction in [3H]-toxin A binding after heat treating cells, which is suggestive of protein binding. The reduction in binding varied between individuals. The reduction of [3H]-toxin A binding, after the removal of beta-linked galactose units, implicates these as components of the receptor and adds credence to the idea that the Lewis X, Y and I antigens may be involved in toxin A binding to human intestinal epithelial cells. However, because the Lewis antigens do not possess terminal alpha-galactose units, the reduction in binding after alpha-galactosidase treatment suggests that other receptors may be involved in toxin A binding to some human intestinal cells. These data are the first demonstration of direct toxin A binding to human intestinal epithelial cells.
authors.affiliation: Medical Research Centre, City Hospital, Nottingham.
authors.forename: J A
authors.initials: JA
authors.lastname: Smith
authors.forename: D L
authors.initials: DL
authors.lastname: Cooke
authors.forename: S
authors.initials: S
authors.lastname: Hyde
authors.forename: S P
authors.initials: SP
authors.lastname: Borriello
authors.forename: R G
authors.initials: RG
authors.lastname: Long
mesh_terms: D001427:Bacterial Toxins; D016360:Clostridioides difficile / Q000472:pathogenicity*; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D004386:Duodenum / Q000166:cytology / Q000378:metabolism*; D004768:Enterotoxins / Q000302:isolation & purification / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D006361:Heating; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000378:metabolism; D016256:Lewis X Antigen / Q000378:metabolism; D011485:Protein Binding; D000519:alpha-Galactosidase / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen; C041348:tcdA protein, Clostridium difficile; D000519:alpha-Galactosidase; D001616:beta-Galactosidase; D005690:Galactose