Abstract
Enterococcus faecalis is among the most important nosocomial pathogens. The intestinal mucosa is considered to be the main site used by these bacteria for entrance and dissemination. A better understanding of the mechanisms involved in colonization and invasion of enterococci may help to devise methods to prevent infections in hospitalized patients. Glycosaminoglycans, which are present on the surface of all eukaryotic cells, were investigated with regard to their role as host receptors for adhesion of E. faecalis. Competitive binding assays, enzymatic digestion, and reduction of the sulfation of the glycosaminoglycan chains indicated that heparin and heparan sulfate, but not chondroitin sulfate B, played important roles in adhesion of E. faecalis 12030 to Caco2 cells. By using proteinases and carbohydrate oxidation by sodium meta-periodate to modify the bacterial surface, it could be demonstrated that a sugar-containing molecule rather than a protein is the bacterial ligand mediating adhesion to eukaryotic cells. Preincubation of Caco2 cells with the enterococcal glycolipid diglucosyldiacylglycerol but not other carbohydrate cell wall components inhibited bacterial binding. These results may indicate that heparin and/or heparan sulfate on host epithelial cells and diglucosyldiacylglycerol, either itself or as a partial structure of lipoteichoic acid, are involved in enterococcal adhesion to colonic epithelia, the first step in translocation from the intestinal tract.
Highlights
Ing them to survive in hospital and nursing home settings (5)
In this study we evaluated the role of host glycosaminoglycans and bacterial surface structures in mediating binding of E. faecalis to mammalian cells
Inhibition of the Bacterial Binding to the Colonic Cell Line Caco2 Using Polysulfated Polysaccharides—When confluent monolayers of Caco2 cells were incubated for 2– 4 h at 37 °C with ϳ100 bacteria/cell, adherence of bacteria correlated with the time of incubation
Summary
Cell Culture—A human cell line, Caco, derived from colon carcinoma was used in this study. Cells were cultivated in DMEM supplemented with 10% fetal bovine serum and 1% nonessential amino acids in a 5% CO2 atmosphere. All the experiments were performed on cells between the 15th and 25th passage. Bacterial Strains—The following strains were used in this study: E. faecalis 12030 (13); FA2-2 (14); type 5/type 9 (15);
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