Mucin isolated from rabbit colon inhibits in vitro binding of Escherichia coli RDEC-1.
The rabbit enteric pathogen Escherichia coli RDEC-1 (serotype O15:H-) mediates attaching and effacing binding to colonic epithelium in a manner morphologically identical to that observed in both human enteropathogenic E. coli and enterohemorrhagic E. coli infections. The aim of this study was to determine if colonic mucus and its constituents, including mucin derived from goblet cells, inhibited RDEC-1 adherence in vitro. Crude mucus was prepared from mucosal scrapings of rabbit colon and separated by buoyant density into eight fractions. Purified mucin was characterized by gel electrophoresis, dot immunoblotting, indirect immunofluorescence, and amino acid composition. RDEC-1 bacteria were grown to promote and suppress the expression of mannose-resistant, hydrophobic pili. A nonpiliated mutant, strain M34, was also used as a negative control. Binding of radiolabeled RDEC-1 expressing pili was quantitated in the presence of crude mucus, purified mucin, and nonmucin fractions. Binding of piliated RDEC-1 to hydrophobic polystyrene wells was greater than for both nonpiliated RDEC-1 and strain M34 (P less than 0.05). Both crude mucus and purified mucin mediated a concentration-dependent inhibition of piliated-RDEC-1 binding. Fractions of mucus without immunoreactive mucin did not inhibit the binding of RDEC-1 expressing hydrophobic pili. We conclude that colonic goblet cell-derived mucin mediates inhibition of piliated RDEC-1 attachment in vitro. Inhibition of bacterial adherence could prevent access of attaching and effacing E. coli enteric pathogens to the colonic mucosa in vivo.
Mack
D R
DR
Department of Pediatrics, Hospital for Sick Children, University of Toronto, Canada.
Sherman
P M
PM
D000704:Analysis of Variance; D000818:Animals; D001422:Bacterial Adhesion; D003106:Colon / Q000382:microbiology* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000502:physiology*; D010861:Fimbriae, Bacterial / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D015151:Immunoblotting; D066298:In Vitro Techniques; D008297:Male; D009077:Mucins / Q000302:isolation & purification / Q000502:physiology*; D009093:Mucus / Q000502:physiology; D011817:Rabbits
D009077:Mucins