Intestinal mucin inhibits adhesion of human enteropathogenic Escherichia coli to HEp-2 cells.
Mucins are complex glycoproteins that cover the intestinal mucosal surface and are thought to provide protection against enteropathogenic infections by preventing bacterial adherence and subsequent colonization, invasion and toxin delivery. The aims of this study was to evaluate the capacity of intestinal mucin to inhibit epithelial cell adhesion by human enteropathogenic Escherichia colI (EPEC). EPEC strain E2348/69 (serotype O127:H6), known to produce a 94-kd epithelia cell-adhesion-mediating outer membrane protein called intimin and known to contain an EPEC adherence factor (EAF) plasmid, and three of its mutants were studied. Crude mucus was obtained from rabbit small-ubtestinal mucosal scrapings. Purified mucins were isolated by serial cesium chloride density gradient ultracentrifugation. Bacterial adhesion to HEp-2 epithelial cell monolayers in the presence of purified intestinal mucins was quantified by recovery of live viable bacteria as colony-forming units. EPEC strain E2348/69 adhesion was inhibited in a concentration-dependent manner by increasing amounts of mucin (adhesion suppressed to <1% by 150 microgram mucin). The relative magnitudes of inhibition of adhesion of E2348/69 and its mutants [JPN15 (EAF plasmid negative), JPN15.96(pMAR7) (intimin negative), and JPN15.96 (intimin negative, EAF plasmid negative)] were similar in the presence of equal amounts of mucin. The presence of the intimin protein and EAF plasmid are not necessary to the proficiency of mucin to inhibit HEp-2 epithelial cell adhesion.
Smith
C J
CJ
Department of Pediatrics, University of Nebraska Medical Center, Omaha 68198-5160, USA.
Kaper
J B
JB
Mack
D R
DR
D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects*; D002294:Carcinoma, Squamous Cell / Q000382:microbiology*; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000502:physiology*; D006801:Humans; D007421:Intestine, Small / Q000737:chemistry*; D007822:Laryngeal Neoplasms / Q000382:microbiology*; D008297:Male; D009077:Mucins / Q000302:isolation & purification / Q000494:pharmacology*; D011817:Rabbits; D014407:Tumor Cells, Cultured
D009077:Mucins