Pilus-mediated binding of bovine enterotoxigenic Escherichia coli to calf small intestinal mucins

Mouricout, Michèle; Julien, Raymond

doi:10.1128/iai.55.5.1216-1223.1987

Cited by 58 publications

(22 citation statements)

References 29 publications

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“…Mouricout and Julien demonstrated binding of an enterotoxigenic E. coli strain expressing K99 pilus antigen to mucus prepared from calf small intestine (19). Similar to our findings, binding of bacteria to calf intestinal mucus was mediated by pili (19).…”

Section: Discussionsupporting

confidence: 89%

Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro

1988

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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 microvilius 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 adhesins, 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.

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Section: Discussionsupporting

confidence: 89%

Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro

1988

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“…The assay adopted routinely in this study for monitoring attachment of ETEC to intestinal preparations was based on the ability of pilus-positive bacteria to capture receptor-containing material. This assay structure has several advantages over the immobilisation of cells reported by other investigators [33,34,42]. Adhesive properties of several different strains may be monitored with a single detection antibody.…”

Section: Discussionmentioning

confidence: 99%

Detection of attachment of enterotoxigenicEscherichia coli(ETEC) to human small intestinal cells by enzyme immunoassay

Mynott

Luke

Chandler

1995

FEMS Immunology &Amp; Medical Microbiology

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Simple immunoassays were developed to study the binding between enterocytes of the small intestine and other cell types, and enterotoxigenic Escherichia coli (ETEC). CFA/I or CFA/II pilus protein or CFA-positive E. coli bacteria were immobilised in wells of microtitre plates and incubated with vesicles or crude mucus prepared from human brush border enterocytes. Binding of the cell preparations was detected by adding specific rabbit anti-brush border IgG followed by urease-labelled goat anti-rabbit IgG and urea substrate. The binding of purified CFA/I to human or rabbit small intestine, human oral epithelial cells or Caco-2 cells was detected with specific anti-CFA/I IgG. Both human brush border and mucus-derived preparations were able to attach to ETEC. The binding was CFA-specific and strong enough to withstand several washings. In contrast, CFA/I did not bind to small intestinal cells of non-human small intestinal origin, indicating that there may be important differences in affinity between receptors present on human small intestinal cells and cells of non-human small intestinal origin. Antibodies directed against human small intestinal and non-small intestinal cells did not cross-react with either preparation, indicating that receptors between these different cell sources are different. The EIA proved useful during the identification of a newly-recognised 15 kDa bacterial surface component of ETEC strain H10407P, which may function as a putative attachment factor. The EIAs developed in this study were easy to perform and multiple tests could be performed on small samples, including biopsy samples obtained during endoscopy.

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“…involved in attachment (and colonization) have been shown to be readily inactivated by proteolytic enzymes, including those proteases that are normally active in the small intestine (23,27,30). This was recently investigated in experiments observing the instability of K88 receptor within pig intestines (3a).…”

Section: Discussionmentioning

confidence: 99%

Efficacy of enteric-coated protease in preventing attachment of enterotoxigenic Escherichia coli and diarrheal disease in the RITARD model

1991

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In this study, we report on a novel approach based on modification of the intestinal surface to prevent diarrhea caused by enterotoxigenic Escherichia coli (ETEC). The removable intestinal tie adult rabbit diarrhea (RITARD) model was used to test the efficacy of an enteric-coated protease preparation (Detach; Enzacor Technology Pty. Ltd.) in the prevention of bacterial attachment and diarrheal disease caused by colonization factor antigen I-positive (CFA/I+) E. coli H10407. Protease was administered orally to rabbits 18 h prior to challenge with 10" bacteria. Four groups of rabbits were inoculated with different ETEC strains which produced different combinations of adhesin and enterotoxin or with sterile phosphate-buffered saline. Occurrence of diarrhea during the subsequent 24-h incubation period was recorded. Oral administration of protease was successful in reducing diarrhea and diarrhea-induced death in six of seven (86%) rabbits infected with CFA/I+, heat-stable and heat-labile toxin-positive E. coli (H10407). Seven of eight (87%) rabbits not protected by protease treatment died or developed severe diarrhea. Quantitative analysis of bacterial cultures obtained from the small intestine of rabbits showed a significant (P < 0.001) 2,000-fold reduction in CFU per centimeter of intestine following treatment with protease. The efficacy of protease treatment was 99.5%, with very wide confidence limits (>0 to 99.9%). The data indicate that the use of protease to prevent ETEC diarrheal disease has considerable potential.

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Pilus-mediated binding of bovine enterotoxigenic Escherichia coli to calf small intestinal mucins

Cited by 58 publications

References 29 publications

Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro

Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro

Detection of attachment of enterotoxigenicEscherichia coli(ETEC) to human small intestinal cells by enzyme immunoassay

Efficacy of enteric-coated protease in preventing attachment of enterotoxigenic Escherichia coli and diarrheal disease in the RITARD model

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