Effects of fibronectin and other salivary macromolecules on the adherence of Escherichia coli to buccal epithelial cells.
The effect of saliva and fibronectin (Fn) on the adherence of a type 1 fimbriated strain of Escherichia coli to human buccal epithelial cells was studied. Saliva pretreatment of epithelial cells led to a dose-dependent increase in adherence that was inhibited by alpha-methyl mannoside, which is typical of a type 1 fimbria-mediated event. The molecules responsible for affecting this increased adherence were nondialyzable and were recovered after lyophilization. E. coli adherence was stimulated by individual saliva samples from each of 11 volunteers. Fn inhibited E. coli adherence to saliva-treated buccal cells by more than 60%. Biotinylated E. coli and Fn were reacted with Western blots of whole saliva to identify the receptors that might explain the phenomenon described above. Both E. coli and Fn bound to 57- and 62-kilodalton (kDa) protein bands in Western blots of sodium dodecyl sulfate gels of whole saliva. The binding of E. coli to these bands was inhibited by pretreatment with unlabeled Fn. To study these salivary components, samples of saliva were electrophoresed on sodium dodecyl sulfate gels, strips corresponding to the appropriate molecular weights were cut out, and the proteins were eluted electrophoretically. Material that eluted from strips at 57 and 62 kDa, but not that from a control strip, stimulated E. coli adherence to buccal cells. Alternatively, saliva was fractionated over 100- and 50-kDa cutoff filters. Of the three fractions obtained, only the fraction passing through the 100-kDa filter and retained by the 50-kDa filter stimulated E. coli adherence to buccal cells. This fraction also increased the binding of Fn to buccal cells. These observations suggest the possibility that one or more salivary components bind to the surface of buccal cells and serve as receptors for type 1 fimbriated E. coli. Fn also binds to this isolated material; and it is apparently by these interactions, at least in part, that saliva stimulates and Fn inhibits E. coli adherence. The way in which these interactions may affect bacterial adherence in vivo remains to be elucidated.
Hasty
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Simpson
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D001422:Bacterial Adhesion; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000382:microbiology; D004926:Escherichia coli / Q000472:pathogenicity*; D005353:Fibronectins / Q000502:physiology*; D066298:In Vitro Techniques; D008766:Methylmannosides / Q000378:metabolism; D008970:Molecular Weight; D009061:Mouth Mucosa / Q000166:cytology / Q000382:microbiology*; D012471:Salivary Proteins and Peptides / Q000502:physiology*
D005353:Fibronectins; D008766:Methylmannosides; D012471:Salivary Proteins and Peptides