Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jan;64(1):135–145. doi: 10.1128/iai.64.1.135-145.1996

Adherence of Salmonella typhimurium to Caco-2 cells: identification of a glycoconjugate receptor.

K T Giannasca 1, P J Giannasca 1, M R Neutra 1
PMCID: PMC173738  PMID: 8557331

Abstract

The mechanism by which Salmonella species adhere to the epithelium of the intestine is not well understood. To identify components on intestinal epithelial cells that may be involved in the initial adherence of Salmonella typhimurium, we correlated patterns of adherence to well-differentiated Caco-2BBe cell monolayers with expression of brush border membrane components and lectin binding sites. This cloned cell line shows heterogeneous expression of sucrase-isomaltase and most lectin receptors. S. typhimurium adhered to a subpopulation of living or formaldehyde-fixed cells with a high multiplicity (up to 150 bacteria per cell). Bacterial binding to selected cells was not correlated with expression of the brush border hydrolases dipeptidyl-peptidase IV and sucrase-isomaltase or with binding of 10 of the 12 lectins tested. However, binding was correlated with the presence of binding sites for peanut agglutinin (PNA) [specific for Gal beta (1-3) GalNAc] and soybean agglutinin (specific for terminal GalNAc). Preincubation of live and fixed Caco-2BBe monolayers with PNA inhibited bacterial binding, while preincubation with soybean agglutinin did not. Electron microscopic analysis demonstrated that the initial adherence of S. typhimurium to Caco-2 cells in vitro involved peripheral components of the glycocalyx on apical microvilli. These results suggest that a Gal beta (1-3)GalNAc epitope recognized by PNA and located in the glycocalyx is involved in the early recognition events between S. typhimurium and Caco-2 cells and that differences in glycosylation patterns among individual epithelial cells may be a determinant in cell-selective adherence of S. typhimurium.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen A. K., Neuberger A., Sharon N. The purification, composition and specificity of wheat-germ agglutinin. Biochem J. 1973 Jan;131(1):155–162. doi: 10.1042/bj1310155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beaulieu J. F., Quaroni A. Clonal analysis of sucrase-isomaltase expression in the human colon adenocarcinoma Caco-2 cells. Biochem J. 1991 Dec 15;280(Pt 3):599–608. doi: 10.1042/bj2800599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernadac A., Gorvel J. P., Feracci H., Maroux S. Human blood group A-like determinants as marker of the intracellular pools of glycoproteins in secretory and absorbing of A+ rabbit jejunum. Biol Cell. 1984;50(1):31–36. doi: 10.1111/j.1768-322x.1984.tb00252.x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Carter P. B., Collins F. M. The route of enteric infection in normal mice. J Exp Med. 1974 May 1;139(5):1189–1203. doi: 10.1084/jem.139.5.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark M. A., Jepson M. A., Simmons N. L., Hirst B. H. Preferential interaction of Salmonella typhimurium with mouse Peyer's patch M cells. Res Microbiol. 1994 Sep;145(7):543–552. doi: 10.1016/0923-2508(94)90031-0. [DOI] [PubMed] [Google Scholar]
  7. Costa de Beauregard M. A., Pringault E., Robine S., Louvard D. Suppression of villin expression by antisense RNA impairs brush border assembly in polarized epithelial intestinal cells. EMBO J. 1995 Feb 1;14(3):409–421. doi: 10.1002/j.1460-2075.1995.tb07017.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Debray H., Montreuil J. Aleuria aurantia agglutinin. A new isolation procedure and further study of its specificity towards various glycopeptides and oligosaccharides. Carbohydr Res. 1989 Jan 15;185(1):15–26. doi: 10.1016/0008-6215(89)84017-0. [DOI] [PubMed] [Google Scholar]
  9. Falk P., Roth K. A., Gordon J. I. Lectins are sensitive tools for defining the differentiation programs of mouse gut epithelial cell lineages. Am J Physiol. 1994 Jun;266(6 Pt 1):G987–1003. doi: 10.1152/ajpgi.1994.266.6.G987. [DOI] [PubMed] [Google Scholar]
  10. Finlay B. B., Falkow S. Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J Infect Dis. 1990 Nov;162(5):1096–1106. doi: 10.1093/infdis/162.5.1096. [DOI] [PubMed] [Google Scholar]
  11. Finlay B. B., Heffron F., Falkow S. Epithelial cell surfaces induce Salmonella proteins required for bacterial adherence and invasion. Science. 1989 Feb 17;243(4893):940–943. doi: 10.1126/science.2919285. [DOI] [PubMed] [Google Scholar]
  12. Finlay B. B. Molecular and cellular mechanisms of Salmonella pathogenesis. Curr Top Microbiol Immunol. 1994;192:163–185. doi: 10.1007/978-3-642-78624-2_8. [DOI] [PubMed] [Google Scholar]
  13. Francis C. L., Ryan T. A., Jones B. D., Smith S. J., Falkow S. Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria. Nature. 1993 Aug 12;364(6438):639–642. doi: 10.1038/364639a0. [DOI] [PubMed] [Google Scholar]
  14. Giannasca P. J., Giannasca K. T., Falk P., Gordon J. I., Neutra M. R. Regional differences in glycoconjugates of intestinal M cells in mice: potential targets for mucosal vaccines. Am J Physiol. 1994 Dec;267(6 Pt 1):G1108–G1121. doi: 10.1152/ajpgi.1994.267.6.G1108. [DOI] [PubMed] [Google Scholar]
  15. Ginocchio C., Pace J., Galán J. E. Identification and molecular characterization of a Salmonella typhimurium gene involved in triggering the internalization of salmonellae into cultured epithelial cells. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5976–5980. doi: 10.1073/pnas.89.13.5976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hauri H. P., Sterchi E. E., Bienz D., Fransen J. A., Marxer A. Expression and intracellular transport of microvillus membrane hydrolases in human intestinal epithelial cells. J Cell Biol. 1985 Sep;101(3):838–851. doi: 10.1083/jcb.101.3.838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hohmann A. W., Schmidt G., Rowley D. Intestinal colonization and virulence of Salmonella in mice. Infect Immun. 1978 Dec;22(3):763–770. doi: 10.1128/iai.22.3.763-770.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoyer L. L., Roggentin P., Schauer R., Vimr E. R. Purification and properties of cloned Salmonella typhimurium LT2 sialidase with virus-typical kinetic preference for sialyl alpha 2----3 linkages. J Biochem. 1991 Sep;110(3):462–467. doi: 10.1093/oxfordjournals.jbchem.a123603. [DOI] [PubMed] [Google Scholar]
  19. Hultgren S. J., Abraham S., Caparon M., Falk P., St Geme J. W., 3rd, Normark S. Pilus and nonpilus bacterial adhesins: assembly and function in cell recognition. Cell. 1993 Jun 4;73(5):887–901. doi: 10.1016/0092-8674(93)90269-v. [DOI] [PubMed] [Google Scholar]
  20. Ito S. The enteric surface coat on cat intestinal microvilli. J Cell Biol. 1965 Dec;27(3):475–491. doi: 10.1083/jcb.27.3.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jones B. D., Ghori N., Falkow S. Salmonella typhimurium initiates murine infection by penetrating and destroying the specialized epithelial M cells of the Peyer's patches. J Exp Med. 1994 Jul 1;180(1):15–23. doi: 10.1084/jem.180.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kaladas P. M., Kabat E. A., Iglesias J. L., Lis H., Sharon N. Immunochemical studies on the combining site of the D-galactose/N-acetyl-D-galactosamine specific lectin from Erythrina cristagalli seeds. Arch Biochem Biophys. 1982 Sep;217(2):624–637. doi: 10.1016/0003-9861(82)90544-6. [DOI] [PubMed] [Google Scholar]
  23. Karlsson K. A. Animal glycosphingolipids as membrane attachment sites for bacteria. Annu Rev Biochem. 1989;58:309–350. doi: 10.1146/annurev.bi.58.070189.001521. [DOI] [PubMed] [Google Scholar]
  24. Kohbata S., Yokoyama H., Yabuuchi E. Cytopathogenic effect of Salmonella typhi GIFU 10007 on M cells of murine ileal Peyer's patches in ligated ileal loops: an ultrastructural study. Microbiol Immunol. 1986;30(12):1225–1237. doi: 10.1111/j.1348-0421.1986.tb03055.x. [DOI] [PubMed] [Google Scholar]
  25. Lee C. A., Falkow S. The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4304–4308. doi: 10.1073/pnas.87.11.4304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lindstedt R., Larson G., Falk P., Jodal U., Leffler H., Svanborg C. The receptor repertoire defines the host range for attaching Escherichia coli strains that recognize globo-A. Infect Immun. 1991 Mar;59(3):1086–1092. doi: 10.1128/iai.59.3.1086-1092.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Louvard D., Kedinger M., Hauri H. P. The differentiating intestinal epithelial cell: establishment and maintenance of functions through interactions between cellular structures. Annu Rev Cell Biol. 1992;8:157–195. doi: 10.1146/annurev.cb.08.110192.001105. [DOI] [PubMed] [Google Scholar]
  28. Maiuri L., Raia V., Fiocca R., Solcia E., Cornaggia M., Norèn O., Sjostrom H., Swallow D., Auricchio S., Dabelsteen E. Mosaic differentiation of human villus enterocytes: patchy expression of blood group A antigen in A nonsecretors. Gastroenterology. 1993 Jan;104(1):21–30. doi: 10.1016/0016-5085(93)90831-v. [DOI] [PubMed] [Google Scholar]
  29. Maury J., Nicoletti C., Guzzo-Chambraud L., Maroux S. The filamentous brush border glycocalyx, a mucin-like marker of enterocyte hyper-polarization. Eur J Biochem. 1995 Mar 1;228(2):323–331. [PubMed] [Google Scholar]
  30. Miller R. L. Properties of a sialic acid-specific lectin from the slug Limax flavus. Methods Enzymol. 1987;138:527–536. doi: 10.1016/0076-6879(87)38047-4. [DOI] [PubMed] [Google Scholar]
  31. Monsigny M., Roche A. C., Sene C., Maget-Dana R., Delmotte F. Sugar-lectin interactions: how does wheat-germ agglutinin bind sialoglycoconjugates? Eur J Biochem. 1980 Feb;104(1):147–153. doi: 10.1111/j.1432-1033.1980.tb04410.x. [DOI] [PubMed] [Google Scholar]
  32. Mooseker M. S. Organization, chemistry, and assembly of the cytoskeletal apparatus of the intestinal brush border. Annu Rev Cell Biol. 1985;1:209–241. doi: 10.1146/annurev.cb.01.110185.001233. [DOI] [PubMed] [Google Scholar]
  33. Müller K. H., Collinson S. K., Trust T. J., Kay W. W. Type 1 fimbriae of Salmonella enteritidis. J Bacteriol. 1991 Aug;173(15):4765–4772. doi: 10.1128/jb.173.15.4765-4772.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nicolson G. L., Blaustein J., Etzler M. E. Characterization of two plant lectins from Ricinus communis and their quantitative interaction with a murine lymphoma. Biochemistry. 1974 Jan 1;13(1):196–204. doi: 10.1021/bi00698a029. [DOI] [PubMed] [Google Scholar]
  35. Pereira M. E., Kisailus E. C., Gruezo F., Kabat E. A. Immunochemical studies on the combining site of the blood group H-specific lectin 1 from Ulex europeus seeds. Arch Biochem Biophys. 1978 Jan 15;185(1):108–115. doi: 10.1016/0003-9861(78)90149-2. [DOI] [PubMed] [Google Scholar]
  36. Peterson M. D., Mooseker M. S. Characterization of the enterocyte-like brush border cytoskeleton of the C2BBe clones of the human intestinal cell line, Caco-2. J Cell Sci. 1992 Jul;102(Pt 3):581–600. doi: 10.1242/jcs.102.3.581. [DOI] [PubMed] [Google Scholar]
  37. Phillips T. E., Huet C., Bilbo P. R., Podolsky D. K., Louvard D., Neutra M. R. Human intestinal goblet cells in monolayer culture: characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line. Gastroenterology. 1988 Jun;94(6):1390–1403. doi: 10.1016/0016-5085(88)90678-6. [DOI] [PubMed] [Google Scholar]
  38. Piller V., Piller F., Cartron J. P. Comparison of the carbohydrate-binding specificities of seven N-acetyl-D-galactosamine-recognizing lectins. Eur J Biochem. 1990 Jul 31;191(2):461–466. doi: 10.1111/j.1432-1033.1990.tb19144.x. [DOI] [PubMed] [Google Scholar]
  39. Quaroni A., Isselbacher K. J. Study of intestinal cell differentiation with monoclonal antibodies to intestinal cell surface components. Dev Biol. 1985 Oct;111(2):267–279. doi: 10.1016/0012-1606(85)90482-8. [DOI] [PubMed] [Google Scholar]
  40. Roberts J. A., Marklund B. I., Ilver D., Haslam D., Kaack M. B., Baskin G., Louis M., Möllby R., Winberg J., Normark S. The Gal(alpha 1-4)Gal-specific tip adhesin of Escherichia coli P-fimbriae is needed for pyelonephritis to occur in the normal urinary tract. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11889–11893. doi: 10.1073/pnas.91.25.11889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Saiman L., Prince A. Pseudomonas aeruginosa pili bind to asialoGM1 which is increased on the surface of cystic fibrosis epithelial cells. J Clin Invest. 1993 Oct;92(4):1875–1880. doi: 10.1172/JCI116779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sastry M. V., Banarjee P., Patanjali S. R., Swamy M. J., Swarnalatha G. V., Surolia A. Analysis of saccharide binding to Artocarpus integrifolia lectin reveals specific recognition of T-antigen (beta-D-Gal(1----3)D-GalNAc). J Biol Chem. 1986 Sep 5;261(25):11726–11733. [PubMed] [Google Scholar]
  43. Semenza G. Anchoring and biosynthesis of stalked brush border membrane proteins: glycosidases and peptidases of enterocytes and renal tubuli. Annu Rev Cell Biol. 1986;2:255–313. doi: 10.1146/annurev.cb.02.110186.001351. [DOI] [PubMed] [Google Scholar]
  44. Sharon N. Bacterial lectins, cell-cell recognition and infectious disease. FEBS Lett. 1987 Jun 15;217(2):145–157. doi: 10.1016/0014-5793(87)80654-3. [DOI] [PubMed] [Google Scholar]
  45. Swamy M. J., Gupta D., Mahanta S. K., Surolia A. Further characterization of the saccharide specificity of peanut (Arachis hypogaea) agglutinin. Carbohydr Res. 1991 Jun 25;213:59–67. doi: 10.1016/s0008-6215(00)90598-6. [DOI] [PubMed] [Google Scholar]
  46. Takeuchi A. Electron microscope studies of experimental Salmonella infection. I. Penetration into the intestinal epithelium by Salmonella typhimurium. Am J Pathol. 1967 Jan;50(1):109–136. [PMC free article] [PubMed] [Google Scholar]
  47. Tollefsen S. E., Kornfeld R. Isolation and characterization of lectins from Vicia villosa. Two distinct carbohydrate binding activities are present in seed extracts. J Biol Chem. 1983 Apr 25;258(8):5165–5171. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES