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. 1997 Feb;65(2):750–757. doi: 10.1128/iai.65.2.750-757.1997

Inhibition of Helicobacter pylori binding to gastrointestinal epithelial cells by sialic acid-containing oligosaccharides.

P M Simon 1, P L Goode 1, A Mobasseri 1, D Zopf 1
PMCID: PMC176121  PMID: 9009338

Abstract

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.

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Selected References

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  1. Ascencio F., Fransson L. A., Wadström T. Affinity of the gastric pathogen Helicobacter pylori for the N-sulphated glycosaminoglycan heparan sulphate. J Med Microbiol. 1993 Apr;38(4):240–244. doi: 10.1099/00222615-38-4-240. [DOI] [PubMed] [Google Scholar]
  2. Bezouska K., Yuen C. T., O'Brien J., Childs R. A., Chai W., Lawson A. M., Drbal K., Fiserová A., Pospísil M., Feizi T. Oligosaccharide ligands for NKR-P1 protein activate NK cells and cytotoxicity. Nature. 1994 Nov 10;372(6502):150–157. doi: 10.1038/372150a0. [DOI] [PubMed] [Google Scholar]
  3. Blaser M. J. Helicobacter pylori phenotypes associated with peptic ulceration. Scand J Gastroenterol Suppl. 1994;205:1–5. [PubMed] [Google Scholar]
  4. Borhan-Manesh F., Farnum J. B. Study of Helicobacter pylori colonization of patches of heterotopic gastric mucosa (HGM) at the upper esophagus. Dig Dis Sci. 1993 Jan;38(1):142–146. doi: 10.1007/BF01296787. [DOI] [PubMed] [Google Scholar]
  5. Borén T., Falk P., Roth K. A., Larson G., Normark S. Attachment of Helicobacter pylori to human gastric epithelium mediated by blood group antigens. Science. 1993 Dec 17;262(5141):1892–1895. doi: 10.1126/science.8018146. [DOI] [PubMed] [Google Scholar]
  6. Chmiela M., Paziak-Domanska B., Rudnicka W., Wadström T. The role of heparan sulphate-binding activity of Helicobacter pylori bacteria in their adhesion to murine macrophages. APMIS. 1995 Jun;103(6):469–474. doi: 10.1111/j.1699-0463.1995.tb01133.x. [DOI] [PubMed] [Google Scholar]
  7. Dunn B. E., Altmann M., Campbell G. P. Adherence of Helicobacter pylori to gastric carcinoma cells: analysis by flow cytometry. Rev Infect Dis. 1991 Jul-Aug;13 (Suppl 8):S657–S664. doi: 10.1093/clinids/13.supplement_8.s657. [DOI] [PubMed] [Google Scholar]
  8. Dwarakanath A. D., Tsai H. H., Sunderland D., Hart C. A., Figura N., Crabtree J. E., Rhodes J. M. The production of neuraminidase and fucosidase by Helicobacter pylori: their possible relationship to pathogenicity. FEMS Immunol Med Microbiol. 1995 Dec;12(3-4):213–216. doi: 10.1111/j.1574-695X.1995.tb00194.x. [DOI] [PubMed] [Google Scholar]
  9. Eaton K. A., Brooks C. L., Morgan D. R., Krakowka S. Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun. 1991 Jul;59(7):2470–2475. doi: 10.1128/iai.59.7.2470-2475.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eaton K. A., Catrenich C. E., Makin K. M., Krakowka S. Virulence of coccoid and bacillary forms of Helicobacter pylori in gnotobiotic piglets. J Infect Dis. 1995 Feb;171(2):459–462. doi: 10.1093/infdis/171.2.459. [DOI] [PubMed] [Google Scholar]
  11. Erbe D. V., Watson S. R., Presta L. G., Wolitzky B. A., Foxall C., Brandley B. K., Lasky L. A. P- and E-selectin use common sites for carbohydrate ligand recognition and cell adhesion. J Cell Biol. 1993 Mar;120(5):1227–1235. doi: 10.1083/jcb.120.5.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Evans D. G., Evans D. J., Jr Adhesion properties of Helicobacter pylori. Methods Enzymol. 1995;253:336–360. doi: 10.1016/s0076-6879(95)53029-0. [DOI] [PubMed] [Google Scholar]
  13. Evans D. G., Evans D. J., Jr, Graham D. Y. Receptor-mediated adherence of Campylobacter pylori to mouse Y-1 adrenal cell monolayers. Infect Immun. 1989 Aug;57(8):2272–2278. doi: 10.1128/iai.57.8.2272-2278.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Evans D. G., Evans D. J., Jr, Lampert H. C., Graham D. Y. Restriction fragment length polymorphism in the adhesin gene hpaA of Helicobacter pylori. Am J Gastroenterol. 1995 Aug;90(8):1282–1288. [PubMed] [Google Scholar]
  15. Evans D. G., Evans D. J., Jr, Moulds J. J., Graham D. Y. N-acetylneuraminyllactose-binding fibrillar hemagglutinin of Campylobacter pylori: a putative colonization factor antigen. Infect Immun. 1988 Nov;56(11):2896–2906. doi: 10.1128/iai.56.11.2896-2906.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Evans D. G., Karjalainen T. K., Evans D. J., Jr, Graham D. Y., Lee C. H. Cloning, nucleotide sequence, and expression of a gene encoding an adhesin subunit protein of Helicobacter pylori. J Bacteriol. 1993 Feb;175(3):674–683. doi: 10.1128/jb.175.3.674-683.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Falk P., Roth K. A., Borén T., Westblom T. U., Gordon J. I., Normark S. An in vitro adherence assay reveals that Helicobacter pylori exhibits cell lineage-specific tropism in the human gastric epithelium. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):2035–2039. doi: 10.1073/pnas.90.5.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fauchère J. L., Blaser M. J. Adherence of Helicobacter pylori cells and their surface components to HeLa cell membranes. Microb Pathog. 1990 Dec;9(6):427–439. doi: 10.1016/0882-4010(90)90061-t. [DOI] [PubMed] [Google Scholar]
  19. Finkelstein R. A., Burks M. F., Zupan A., Dallas W. S., Jacob C. O., Ludwig D. S. Epitopes of the cholera family of enterotoxins. Rev Infect Dis. 1987 May-Jun;9(3):544–561. doi: 10.1093/clinids/9.3.544. [DOI] [PubMed] [Google Scholar]
  20. Fu D., van Halbeek H. N-glycosylation site mapping of human serotransferrin by serial lectin affinity chromatography, fast atom bombardment-mass spectrometry, and 1H nuclear magnetic resonance spectroscopy. Anal Biochem. 1992 Oct;206(1):53–63. doi: 10.1016/s0003-2697(05)80010-7. [DOI] [PubMed] [Google Scholar]
  21. Goldhar J. Erythrocytes as target cells for testing bacterial adhesins. Methods Enzymol. 1995;253:43–50. doi: 10.1016/s0076-6879(95)53006-1. [DOI] [PubMed] [Google Scholar]
  22. Hansson G. C., Karlsson K. A., Larson G., Strömberg N., Thurin J., Orvell C., Norrby E. A novel approach to the study of glycolipid receptors for viruses. Binding of Sendai virus to thin-layer chromatograms. FEBS Lett. 1984 May 7;170(1):15–18. doi: 10.1016/0014-5793(84)81359-9. [DOI] [PubMed] [Google Scholar]
  23. Hoogewerf A. J., Cisar L. A., Evans D. C., Bensadoun A. Effect of chlorate on the sulfation of lipoprotein lipase and heparan sulfate proteoglycans. Sulfation of heparan sulfate proteoglycans affects lipoprotein lipase degradation. J Biol Chem. 1991 Sep 5;266(25):16564–16571. [PubMed] [Google Scholar]
  24. Huang J., Keeling P. W., Smyth C. J. Identification of erythrocyte-binding antigens in Helicobacter pylori. J Gen Microbiol. 1992 Jul;138(7):1503–1513. doi: 10.1099/00221287-138-7-1503. [DOI] [PubMed] [Google Scholar]
  25. Hurtado A., Chahal B., Owen R. J., Smith A. W. Genetic diversity of the Helicobacter pylori haemagglutinin/protease (hap) gene. FEMS Microbiol Lett. 1994 Oct 15;123(1-2):173–178. doi: 10.1111/j.1574-6968.1994.tb07218.x. [DOI] [PubMed] [Google Scholar]
  26. Hussell T., Isaacson P. G., Crabtree J. E., Spencer J. The response of cells from low-grade B-cell gastric lymphomas of mucosa-associated lymphoid tissue to Helicobacter pylori. Lancet. 1993 Sep 4;342(8871):571–574. doi: 10.1016/0140-6736(93)91408-e. [DOI] [PubMed] [Google Scholar]
  27. Kamisago S., Iwamori M., Tai T., Mitamura K., Yazaki Y., Sugano K. Role of sulfatides in adhesion of Helicobacter pylori to gastric cancer cells. Infect Immun. 1996 Feb;64(2):624–628. doi: 10.1128/iai.64.2.624-628.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Karjalainen T. K., Evans D. G., So M., Lee C. H. Molecular cloning and nucleotide sequence of the colonization factor antigen I gene of Escherichia coli. Infect Immun. 1989 Apr;57(4):1126–1130. doi: 10.1128/iai.57.4.1126-1130.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Karlsson K. A., Angström J., Bergström J., Lanne B. Microbial interaction with animal cell surface carbohydrates. APMIS Suppl. 1992;27:71–83. [PubMed] [Google Scholar]
  30. Kobayashi Y., Okazaki K., Murakami K. Adhesion of Helicobacter pylori to gastric epithelial cells in primary cultures obtained from stomachs of various animals. Infect Immun. 1993 Oct;61(10):4058–4063. doi: 10.1128/iai.61.10.4058-4063.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Krivan H. C., Roberts D. D., Ginsburg V. Many pulmonary pathogenic bacteria bind specifically to the carbohydrate sequence GalNAc beta 1-4Gal found in some glycolipids. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6157–6161. doi: 10.1073/pnas.85.16.6157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. LIPKIN M., SHERLOCK P., BELL B. CELL PROLIFERATION KINETICS IN THE GASTROINTESTINAL TRACT OF MAN. II. CELL RENEWAL IN STOMACH, ILEUM, COLON, AND RECTUM. Gastroenterology. 1963 Dec;45:721–729. [PubMed] [Google Scholar]
  33. Lee A. H pylori-initiated ulcerogenesis: look to the host. Lancet. 1993 Jan 30;341(8840):280–281. doi: 10.1016/0140-6736(93)92624-3. [DOI] [PubMed] [Google Scholar]
  34. Lingwood C. A., Huesca M., Kuksis A. The glycerolipid receptor for Helicobacter pylori (and exoenzyme S) is phosphatidylethanolamine. Infect Immun. 1992 Jun;60(6):2470–2474. doi: 10.1128/iai.60.6.2470-2474.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Neman-Simha V., Mégraud F. In vitro model for Campylobacter pylori adherence properties. Infect Immun. 1988 Dec;56(12):3329–3333. doi: 10.1128/iai.56.12.3329-3333.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. O'Toole P. W., Janzon L., Doig P., Huang J., Kostrzynska M., Trust T. J. The putative neuraminyllactose-binding hemagglutinin HpaA of Helicobacter pylori CCUG 17874 is a lipoprotein. J Bacteriol. 1995 Nov;177(21):6049–6057. doi: 10.1128/jb.177.21.6049-6057.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Parkkinen J., Finne J. Isolation of sialyl oligosaccharides and sialyl oligosaccharide phosphates from bovine colostrum and human urine. Methods Enzymol. 1987;138:289–300. doi: 10.1016/0076-6879(87)38024-3. [DOI] [PubMed] [Google Scholar]
  38. Parkkinen J., Rogers G. N., Korhonen T., Dahr W., Finne J. Identification of the O-linked sialyloligosaccharides of glycophorin A as the erythrocyte receptors for S-fimbriated Escherichia coli. Infect Immun. 1986 Oct;54(1):37–42. doi: 10.1128/iai.54.1.37-42.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Piotrowski J., Slomiany A., Murty V. L., Fekete Z., Slomiany B. L. Inhibition of Helicobacter pylori colonization by sulfated gastric mucin. Biochem Int. 1991 Jul;24(4):749–756. [PubMed] [Google Scholar]
  40. Pritchett T. J., Paulson J. C. Basis for the potent inhibition of influenza virus infection by equine and guinea pig alpha 2-macroglobulin. J Biol Chem. 1989 Jun 15;264(17):9850–9858. [PubMed] [Google Scholar]
  41. Pérez-Pérez G. I., Olivares A. Z., Cover T. L., Blaser M. J. Characteristics of Helicobacter pylori variants selected for urease deficiency. Infect Immun. 1992 Sep;60(9):3658–3663. doi: 10.1128/iai.60.9.3658-3663.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Saitoh T., Natomi H., Zhao W. L., Okuzumi K., Sugano K., Iwamori M., Nagai Y. Identification of glycolipid receptors for Helicobacter pylori by TLC-immunostaining. FEBS Lett. 1991 May 6;282(2):385–387. doi: 10.1016/0014-5793(91)80519-9. [DOI] [PubMed] [Google Scholar]
  43. Slomiany B. L., Piotrowski J., Samanta A., VanHorn K., Murty V. L., Slomiany A. Campylobacter pylori colonization factor shows specificity for lactosylceramide sulfate and GM3 ganglioside. Biochem Int. 1989 Oct;19(4):929–936. [PubMed] [Google Scholar]
  44. Talley N. J., Zinsmeister A. R., Weaver A., DiMagno E. P., Carpenter H. A., Perez-Perez G. I., Blaser M. J. Gastric adenocarcinoma and Helicobacter pylori infection. J Natl Cancer Inst. 1991 Dec 4;83(23):1734–1739. doi: 10.1093/jnci/83.23.1734. [DOI] [PubMed] [Google Scholar]
  45. Trust T. J., Doig P., Emödy L., Kienle Z., Wadström T., O'Toole P. High-affinity binding of the basement membrane proteins collagen type IV and laminin to the gastric pathogen Helicobacter pylori. Infect Immun. 1991 Dec;59(12):4398–4404. doi: 10.1128/iai.59.12.4398-4404.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tsuda M., Karita M., Morshed M. G., Okita K., Nakazawa T. A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. Infect Immun. 1994 Aug;62(8):3586–3589. doi: 10.1128/iai.62.8.3586-3589.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tzouvelekis L. S., Mentis A. F., Makris A. M., Spiliadis C., Blackwell C., Weir D. M. In vitro binding of Helicobacter pylori to human gastric mucin. Infect Immun. 1991 Nov;59(11):4252–4254. doi: 10.1128/iai.59.11.4252-4254.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Varki A. Selectin ligands. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7390–7397. doi: 10.1073/pnas.91.16.7390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Welply J. K., Abbas S. Z., Scudder P., Keene J. L., Broschat K., Casnocha S., Gorka C., Steininger C., Howard S. C., Schmuke J. J. Multivalent sialyl-LeX: potent inhibitors of E-selectin-mediated cell adhesion; reagent for staining activated endothelial cells. Glycobiology. 1994 Jun;4(3):259–265. doi: 10.1093/glycob/4.3.259. [DOI] [PubMed] [Google Scholar]
  50. Wotherspoon A. C., Doglioni C., Diss T. C., Pan L., Moschini A., de Boni M., Isaacson P. G. Regression of primary low-grade B-cell gastric lymphoma of mucosa-associated lymphoid tissue type after eradication of Helicobacter pylori. Lancet. 1993 Sep 4;342(8871):575–577. doi: 10.1016/0140-6736(93)91409-f. [DOI] [PubMed] [Google Scholar]
  51. Zhang J. P., Normark S. Induction of gene expression in Escherichia coli after pilus-mediated adherence. Science. 1996 Aug 30;273(5279):1234–1236. doi: 10.1126/science.273.5279.1234. [DOI] [PubMed] [Google Scholar]
  52. Zopf D., Roth S. Oligosaccharide anti-infective agents. Lancet. 1996 Apr 13;347(9007):1017–1021. doi: 10.1016/s0140-6736(96)90150-6. [DOI] [PubMed] [Google Scholar]

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