Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro.
Intestinal mucus is postulated to play a role in preventing colonization of the gastrointestinal tract by microbial pathogens. To evaluate the ability of both crude mucus and purified mucin, a glycoprotein of goblet cell origin, to inhibit mucosal adherence of enteric pathogens, we examined whether mucus and mucin derived from rabbit ileum interact with the rabbit enteropathogen Escherichia coli RDEC-1. We examined the manner in which mucus and mucin inhibited adherence of bacteria to rabbit ileal microvillus membranes (MVMs) in vitro. The purity of the mucin preparation was demonstrated by polyacrylamide gel electrophoresis before and after reduction and by showing that an antiserum raised to the mucin localized to goblet cells in rabbit intestine. Using radioactive labeling of bacteria, we quantitated attachment of RDEC-1 to MVMs, mucus, and mucin that had been immobilized on polystyrene microtiter wells. Binding of RDEC-1 to MVMs was also determined after preincubation of organisms with crude ileal mucus and purified mucin. RDEC-1 bound to both crude mucus and purified mucin when they expressed lectinlike adhesions, previously designated attachment factor rabbit 1 pili. Adherence of piliated RDEC-1 to MVMs, mucus, and mucin was significantly greater than when the bacteria were nonpiliated. Binding of piliated RDEC-1 to MVMs was decreased by preincubation of bacteria with both crude mucus (45.6 +/- 4.2% of control) and purified mucin (50.2 +/- 5.8%). These data indicate that the E. coli enteropathogen RDEC-1 can bind to purified glycoproteins of goblet cell origin and that adherence of these bacteria to mucin is mediated by expression of pili. The findings also support a role for intestinal mucus and its principal organic constituent, mucin, in preventing adherence of a known E. coli enteric pathogen to apical MVMs of enterocytes.
D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects; D001667:Binding, Competitive; D004926:Escherichia coli / Q000187:drug effects / Q000502:physiology*; D006168:Guinea Pigs; D007082:Ileum; D007413:Intestinal Mucosa / Q000382:microbiology*; D008297:Male; D008871:Microvilli / Q000382:microbiology / Q000502:physiology; D009077:Mucins / Q000302:isolation & purification / Q000502:physiology*; D009093:Mucus / Q000382:microbiology / Q000502:physiology*; D011817:Rabbits
D009077:Mucins