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Infect. Immun., Feb 1997, 750-757, Vol 65, No. 2
PM Simon, PL Goode, A Mobasseri and D Zopf
Helicobacterpylori, the ulcer pathogen residing in the human stomach, binds
to epithelial cells of the gastric antrum. We have examined binding of 13
bacterial isolates to epithelial cell lines by use of a sensitive
microtiter plate method in which measurement of bacterial urease activity
provides the means for quantitation of bound organisms. Several established
human gastrointestinal carcinoma cell lines grown as monolayers were
compared for suitability in these assays, and the duodenum-derived cell
line HuTu-80 was selected for testing bacterial binding inhibitors. When
bacteria are pretreated with oligosaccharides, glycoproteins, and
glycolipids, a complex picture of bacterial- epithelial adherence
specificities emerges. Among the monovalent inhibitors tested,
3'-sialyllactose (NeuAc alpha2-3Gal beta1-4Glc; 3'SL) was the most active
oligosaccharide, inhibiting adherence for recent clinical isolates of H.
pylori with a millimolar 50% inhibitory concentration (IC50). Its alpha2-6
isomer (6'SL) was less active. Most of the recent clinical isolates
examined were inhibited by sialyllactose, whereas long-passaged isolates
were insensitive. Among the long-passaged bacterial strains whose binding
was not inhibited by 3'SL was the strain ATCC 43504, also known as NCTC
11637 and CCUG 17874, in which the proposed sialyllactose adhesin was
recently reported to lack surface expression (P. G. O'Toole, L. Janzon, P.
Doig, J. Huang, M. Kostrzynska, and T. H. Trust, J. Bacteriol.
177:6049-6057, 1995). Pretreatment of the epithelial monolayer with
neuraminidase reduced the extent of binding by those bacteria that are
sensitive to inhibition by 3'SL. Other potent inhibitors of bacterial
binding are the glycoproteins alpha1-acid glycoprotein, fetuin, porcine
gastric and bovine submaxillary mucins, and the glycolipid sulfatide, all
of which present multivalent sialylated and/or sulfated galactosyl residues
under the conditions of the binding assay. Consistent with this pattern, a
multivalent neoglycoconjugate containing 20 mol of 3'SL per mol of human
serum albumin inhibited bacterial binding with micromolar IC50. The H.
pylori isolate most sensitive to inhibition by 3'SL was least sensitive to
inhibition by sulfatide, gastric mucin, and other sulfated
oligosaccharides. Bacteria that have been allowed to bind epithelial cells
are also effectively detached by 3'SL. These results describe a
heterogeneous adherence repertoire for these bacteria, but they also
confirm the critical role of the 3'SL structure on human gastric epithelial
cells as an adherence ligand for recent isolates of H. pylori.
Copyright © 1997, American Society for Microbiology
Inhibition of Helicobacter pylori binding to gastrointestinal epithelial cells by sialic acid-containing oligosaccharides
Neose Technologies, Inc., Horsham, Pennsylvania 19044, USA. SimonPM@AOL.com
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