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. 1997 Apr;65(4):1299–1306. doi: 10.1128/iai.65.4.1299-1306.1997

Glucose up-regulates expression of the differentiation-associated brush border binding site for enterotoxigenic Escherichia coli colonization factor antigen I in cultured human enterocyte-like cells.

M F Bernet-Camard 1, F Duigou 1, S Kernéis 1, M H Coconnier 1, A L Servin 1
PMCID: PMC175131  PMID: 9119465

Abstract

The association of enterotoxigenic Escherichia coli expressing colonization factor antigen I (CFA/I) with the cultured human colon adenocarcinoma cell, a model of the mature enterocyte of the small intestine, is dependent on the binding of CFA/I to a brush border-associated component. Binding of the purified radiolabeled [125I]CFA/I- and 14C-labeled CFA/I-positive bacteria could be displaced by an increasing concentration of unlabeled CFA/I. Moreover, we showed that expression of the specific CFA/I binding developed as a function of cell differentiation in Caco-2 cells, whereas expression of the nonspecific binding did not. Expression of the brush border differentiation-associated component acting as a binding site for CFA/I was up-regulated by glucose. Indeed, the enterocyte-like HT-29 glc- cell subpopulation not expressing the CFA/I binding site when cultured in dialyzed serum and hexose-free medium regained the ability to bind CFA/I when the cells were returned to culture medium containing glucose. Furthermore, expression of the brush border-associated CFA/I binding site in the enterocyte-like Caco-2 cells was repressed when the cells were cultured in hexose-free conditions.

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Selected References

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  1. Bartus H., Actor P., Snipes E., Sedlock D., Zajac I. Indications that the erythrocyte receptor involved in enterotoxigenic Escherichia coli attachment is a sialoglycoconjugate. J Clin Microbiol. 1985 Jun;21(6):951–954. doi: 10.1128/jcm.21.6.951-954.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernet-Camard M. F., Coconnier M. H., Hudault S., Servin A. L. Pathogenicity of the diffusely adhering strain Escherichia coli C1845: F1845 adhesin-decay accelerating factor interaction, brush border microvillus injury, and actin disassembly in cultured human intestinal epithelial cells. Infect Immun. 1996 Jun;64(6):1918–1928. doi: 10.1128/iai.64.6.1918-1928.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bühler T., Hoschützky H., Jann K. Analysis of colonization factor antigen I, an adhesin of enterotoxigenic Escherichia coli O78:H11: fimbrial morphology and location of the receptor-binding site. Infect Immun. 1991 Nov;59(11):3876–3882. doi: 10.1128/iai.59.11.3876-3882.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chantret I., Lacasa M., Chevalier G., Ruf J., Islam I., Mantei N., Edwards Y., Swallow D., Rousset M. Sequence of the complete cDNA and the 5' structure of the human sucrase-isomaltase gene. Possible homology with a yeast glucoamylase. Biochem J. 1992 Aug 1;285(Pt 3):915–923. doi: 10.1042/bj2850915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chantret I., Rodolosse A., Barbat A., Dussaulx E., Brot-Laroche E., Zweibaum A., Rousset M. Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: evidence for glucose-dependent negative regulation. J Cell Sci. 1994 Jan;107(Pt 1):213–225. doi: 10.1242/jcs.107.1.213. [DOI] [PubMed] [Google Scholar]
  6. Coconnier M. H., Bernet-Camard M. F., Servin A. L. How intestinal epithelial cell differentiation inhibits the cell-entry of Yersinia pseudotuberculosis in colon carcinoma Caco-2 cell line in culture. Differentiation. 1994 Nov;58(1):87–94. doi: 10.1046/j.1432-0436.1994.5810087.x. [DOI] [PubMed] [Google Scholar]
  7. Cohen M. B., Jensen N. J., Hawkins J. A., Mann E. A., Thompson M. R., Lentze M. J., Giannella R. A. Receptors for Escherichia coli heat stable enterotoxin in human intestine and in a human intestinal cell line (Caco-2). J Cell Physiol. 1993 Jul;156(1):138–144. doi: 10.1002/jcp.1041560119. [DOI] [PubMed] [Google Scholar]
  8. Currie M. G., Fok K. F., Kato J., Moore R. J., Hamra F. K., Duffin K. L., Smith C. E. Guanylin: an endogenous activator of intestinal guanylate cyclase. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):947–951. doi: 10.1073/pnas.89.3.947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Danielsen E. M. Folding of intestinal brush border enzymes. Evidence that high-mannose glycosylation is an essential early event. Biochemistry. 1992 Mar 3;31(8):2266–2272. doi: 10.1021/bi00123a008. [DOI] [PubMed] [Google Scholar]
  10. Danielsen E. M. Post-translational suppression of expression of intestinal brush border enzymes by fructose. J Biol Chem. 1989 Aug 15;264(23):13726–13729. [PubMed] [Google Scholar]
  11. Darfeuille-Michaud A., Aubel D., Chauviere G., Rich C., Bourges M., Servin A., Joly B. Adhesion of enterotoxigenic Escherichia coli to the human colon carcinoma cell line Caco-2 in culture. Infect Immun. 1990 Apr;58(4):893–902. doi: 10.1128/iai.58.4.893-902.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dramsi S., Kocks C., Forestier C., Cossart P. Internalin-mediated invasion of epithelial cells by Listeria monocytogenes is regulated by the bacterial growth state, temperature and the pleiotropic activator prfA. Mol Microbiol. 1993 Sep;9(5):931–941. doi: 10.1111/j.1365-2958.1993.tb01223.x. [DOI] [PubMed] [Google Scholar]
  13. Evans D. G., Evans D. J., Jr, Clegg S., Pauley J. A. Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):738–748. doi: 10.1128/iai.25.2.738-748.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eveillard M., Fourel V., Barc M. C., Kernéis S., Coconnier M. H., Karjalainen T., Bourlioux P., Servin A. L. Identification and characterization of adhesive factors of Clostridium difficile involved in adhesion to human colonic enterocyte-like Caco-2 and mucus-secreting HT29 cells in culture. Mol Microbiol. 1993 Feb;7(3):371–381. doi: 10.1111/j.1365-2958.1993.tb01129.x. [DOI] [PubMed] [Google Scholar]
  15. Falkow S., Isberg R. R., Portnoy D. A. The interaction of bacteria with mammalian cells. Annu Rev Cell Biol. 1992;8:333–363. doi: 10.1146/annurev.cb.08.110192.002001. [DOI] [PubMed] [Google Scholar]
  16. Fogh J., Fogh J. M., Orfeo T. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J Natl Cancer Inst. 1977 Jul;59(1):221–226. doi: 10.1093/jnci/59.1.221. [DOI] [PubMed] [Google Scholar]
  17. Forte L. R., Currie M. G. Guanylin: a peptide regulator of epithelial transport. FASEB J. 1995 May;9(8):643–650. doi: 10.1096/fasebj.9.8.7768356. [DOI] [PubMed] [Google Scholar]
  18. Gabastou J. M., Kernéis S., Bernet-Camard M. F., Barbat A., Coconnier M. H., Kaper J. B., Servin A. L. Two stages of enteropathogenic Escherichia coli intestinal pathogenicity are up and down-regulated by the epithelial cell differentiation. Differentiation. 1995 Sep;59(2):127–134. doi: 10.1046/j.1432-0436.1995.5920127.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gaillard J. L., Berche P., Frehel C., Gouin E., Cossart P. Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocci. Cell. 1991 Jun 28;65(7):1127–1141. doi: 10.1016/0092-8674(91)90009-n. [DOI] [PubMed] [Google Scholar]
  20. Gaillard J. L., Berche P., Mounier J., Richard S., Sansonetti P. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun. 1987 Nov;55(11):2822–2829. doi: 10.1128/iai.55.11.2822-2829.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gaillard J. L., Finlay B. B. Effect of cell polarization and differentiation on entry of Listeria monocytogenes into the enterocyte-like Caco-2 cell line. Infect Immun. 1996 Apr;64(4):1299–1308. doi: 10.1128/iai.64.4.1299-1308.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Gyles C. L. Escherichia coli cytotoxins and enterotoxins. Can J Microbiol. 1992 Jul;38(7):734–746. doi: 10.1139/m92-120. [DOI] [PubMed] [Google Scholar]
  23. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  24. Harris D. S., Slot J. W., Geuze H. J., James D. E. Polarized distribution of glucose transporter isoforms in Caco-2 cells. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7556–7560. doi: 10.1073/pnas.89.16.7556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jourdan N., Cotte Laffitte J., Forestier F., Servin A. L., Quéro A. M. Infection of cultured human intestinal cells by monkey RRV and human Wa rotavirus as a function of intestinal epithelial cell differentiation. Res Virol. 1995 Sep-Oct;146(5):325–331. doi: 10.1016/0923-2516(96)80595-4. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Kerneis S., Bernet M. F., Coconnier M. H., Servin A. L. Adhesion of human enterotoxigenic Escherichia coli to human mucus secreting HT-29 cell subpopulations in culture. Gut. 1994 Oct;35(10):1449–1454. doi: 10.1136/gut.35.10.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kerneis S., Bilge S. S., Fourel V., Chauviere G., Coconnier M. H., Servin A. L. Use of purified F1845 fimbrial adhesin to study localization and expression of receptors for diffusely adhering Escherichia coli during enterocytic differentiation of human colon carcinoma cell lines HT-29 and Caco-2 in culture. Infect Immun. 1991 Nov;59(11):4013–4018. doi: 10.1128/iai.59.11.4013-4018.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kernéis S., Chauvière G., Darfeuille-Michaud A., Aubel D., Coconnier M. H., Joly B., Servin A. L. Expression of receptors for enterotoxigenic Escherichia coli during enterocytic differentiation of human polarized intestinal epithelial cells in culture. Infect Immun. 1992 Jul;60(7):2572–2580. doi: 10.1128/iai.60.7.2572-2580.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Knutton S., Lloyd D. R., Candy D. C., McNeish A. S. Ultrastructural study of adhesion of enterotoxigenic Escherichia coli to erythrocytes and human intestinal epithelial cells. Infect Immun. 1984 May;44(2):519–527. doi: 10.1128/iai.44.2.519-527.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis. 1987 Mar;155(3):377–389. doi: 10.1093/infdis/155.3.377. [DOI] [PubMed] [Google Scholar]
  32. Lindahl M., Faris A., Wadstrom T. Colonization factor antigen on enterotoxigenic Escherichia coli is a sialic-specific lectin. Lancet. 1982 Jul 31;2(8292):280–280. doi: 10.1016/s0140-6736(82)90368-3. [DOI] [PubMed] [Google Scholar]
  33. Mahraoui L., Rodolosse A., Barbat A., Dussaulx E., Zweibaum A., Rousset M., Brot-Laroche E. Presence and differential expression of SGLT1, GLUT1, GLUT2, GLUT3 and GLUT5 hexose-transporter mRNAs in Caco-2 cell clones in relation to cell growth and glucose consumption. Biochem J. 1994 Mar 15;298(Pt 3):629–633. doi: 10.1042/bj2980629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mahraoui L., Rousset M., Dussaulx E., Darmoul D., Zweibaum A., Brot-Laroche E. Expression and localization of GLUT-5 in Caco-2 cells, human small intestine, and colon. Am J Physiol. 1992 Sep;263(3 Pt 1):G312–G318. doi: 10.1152/ajpgi.1992.263.3.G312. [DOI] [PubMed] [Google Scholar]
  35. Mann E. A., Cohen M. B., Giannella R. A. Comparison of receptors for Escherichia coli heat-stable enterotoxin: novel receptor present in IEC-6 cells. Am J Physiol. 1993 Jan;264(1 Pt 1):G172–G178. doi: 10.1152/ajpgi.1993.264.1.G172. [DOI] [PubMed] [Google Scholar]
  36. Mengaud J., Ohayon H., Gounon P., Mege R-M, Cossart P. E-cadherin is the receptor for internalin, a surface protein required for entry of L. monocytogenes into epithelial cells. Cell. 1996 Mar 22;84(6):923–932. doi: 10.1016/s0092-8674(00)81070-3. [DOI] [PubMed] [Google Scholar]
  37. Mesonero J., Matosin M., Cambier D., Rodriguez-Yoldi M. J., Brot-Laroche E. Sugar-dependent expression of the fructose transporter GLUT5 in Caco-2 cells. Biochem J. 1995 Dec 15;312(Pt 3):757–762. doi: 10.1042/bj3120757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mounier J., Vasselon T., Hellio R., Lesourd M., Sansonetti P. J. Shigella flexneri enters human colonic Caco-2 epithelial cells through the basolateral pole. Infect Immun. 1992 Jan;60(1):237–248. doi: 10.1128/iai.60.1.237-248.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Neeser J. R., Chambaz A., Golliard M., Link-Amster H., Fryder V., Kolodziejczyk E. Adhesion of colonization factor antigen II-positive enterotoxigenic Escherichia coli strains to human enterocytelike differentiated HT-29 cells: a basis for host-pathogen interactions in the gut. Infect Immun. 1989 Dec;57(12):3727–3734. doi: 10.1128/iai.57.12.3727-3734.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ofek I., Zafriri D., Goldhar J., Eisenstein B. I. Inability of toxin inhibitors to neutralize enhanced toxicity caused by bacteria adherent to tissue culture cells. Infect Immun. 1990 Nov;58(11):3737–3742. doi: 10.1128/iai.58.11.3737-3742.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ogier-Denis E., Bauvy C., Aubery M., Codogno P., Sapin C., Darmoul D., Zweibaum A., Trugnan G. N-glycosylation modification of proteins is an early marker of the enterocytic differentiation process of HT-29 cells. Reprod Nutr Dev. 1990;30(3):325–330. doi: 10.1051/rnd:19900305. [DOI] [PubMed] [Google Scholar]
  42. Ogier-Denis E., Blais A., Houri J. J., Voisin T., Trugnan G., Codogno P. The emergence of a basolateral 1-deoxymannojirimycin-sensitive mannose carrier is a function of intestinal epithelial cell differentiation. Evidence for a new inhibitory effect of 1-deoxymannojirimycin on facilitative mannose transport. J Biol Chem. 1994 Feb 11;269(6):4285–4290. [PubMed] [Google Scholar]
  43. Orlandi P. A., Critchley D. R., Fishman P. H. The heat-labile enterotoxin of Escherichia coli binds to polylactosaminoglycan-containing receptors in CaCo-2 human intestinal epithelial cells. Biochemistry. 1994 Nov 1;33(43):12886–12895. doi: 10.1021/bi00209a021. [DOI] [PubMed] [Google Scholar]
  44. Orø H. S., Kolstø A. B., Wennerås C., Svennerholm A. M. Identification of asialo GM1 as a binding structure for Escherichia coli colonization factor antigens. FEMS Microbiol Lett. 1990 Nov;60(3):289–292. doi: 10.1016/0378-1097(90)90319-l. [DOI] [PubMed] [Google Scholar]
  45. Pieroni P., Worobec E. A., Paranchych W., Armstrong G. D. Identification of a human erythrocyte receptor for colonization factor antigen I pili expressed by H10407 enterotoxigenic Escherichia coli. Infect Immun. 1988 May;56(5):1334–1340. doi: 10.1128/iai.56.5.1334-1340.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Schulz S., Green C. K., Yuen P. S., Garbers D. L. Guanylyl cyclase is a heat-stable enterotoxin receptor. Cell. 1990 Nov 30;63(5):941–948. doi: 10.1016/0092-8674(90)90497-3. [DOI] [PubMed] [Google Scholar]
  47. Spangler B. D. Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin. Microbiol Rev. 1992 Dec;56(4):622–647. doi: 10.1128/mr.56.4.622-647.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Trugnan G., Ogier-Denis E., Sapin C., Darmoul D., Bauvy C., Aubery M., Codogno P. The N-glycan processing in HT-29 cells is a function of their state of enterocytic differentiation. Evidence for an atypical traffic associated with change in polypeptide stability in undifferentiated HT-29 cells. J Biol Chem. 1991 Nov 5;266(31):20849–20855. [PubMed] [Google Scholar]
  49. Visweswariah S. S., Shanthi G., Balganesh T. S. Interaction of heat-stable enterotoxins with human colonic (T84) cells: modulation of the activation of guanylyl cyclase. Microb Pathog. 1992 Mar;12(3):209–218. doi: 10.1016/0882-4010(92)90055-s. [DOI] [PubMed] [Google Scholar]
  50. Wennerås C., Holmgren J., Svennerholm A. M. The binding of colonization factor antigens of enterotoxigenic Escherichia coli to intestinal cell membrane proteins. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):107–112. doi: 10.1016/0378-1097(90)90266-s. [DOI] [PubMed] [Google Scholar]
  51. Wennerås C., Neeser J. R., Svennerholm A. M. Binding of the fibrillar CS3 adhesin of enterotoxigenic Escherichia coli to rabbit intestinal glycoproteins is competitively prevented by GalNAc beta 1-4Gal-containing glycoconjugates. Infect Immun. 1995 Feb;63(2):640–646. doi: 10.1128/iai.63.2.640-646.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zweibaum A., Pinto M., Chevalier G., Dussaulx E., Triadou N., Lacroix B., Haffen K., Brun J. L., Rousset M. Enterocytic differentiation of a subpopulation of the human colon tumor cell line HT-29 selected for growth in sugar-free medium and its inhibition by glucose. J Cell Physiol. 1985 Jan;122(1):21–29. doi: 10.1002/jcp.1041220105. [DOI] [PubMed] [Google Scholar]
  53. de Graaf F. K. Genetics of adhesive fimbriae of intestinal Escherichia coli. Curr Top Microbiol Immunol. 1990;151:29–53. doi: 10.1007/978-3-642-74703-8_2. [DOI] [PubMed] [Google Scholar]

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