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. 1993 Nov 2;123(4):895–907. doi: 10.1083/jcb.123.4.895

Salmonella typhimurium attachment to human intestinal epithelial monolayers: transcellular signalling to subepithelial neutrophils

PMCID: PMC2200157  PMID: 8227148

Abstract

In human intestinal disease induced by Salmonella typhimurium, transepithelial migration of neutrophils (PMN) rapidly follows attachment of the bacteria to the epithelial apical membrane. In this report, we model those interactions in vitro, using polarized monolayers of the human intestinal epithelial cell, T84, isolated human PMN, and S. typhimurium. We show that Salmonella attachment to T84 cell apical membranes did not alter monolayer integrity as assessed by transepithelial resistance and measurements of ion transport. However, when human neutrophils were subsequently placed on the basolateral surface of monolayers apically colonized by Salmonella, physiologically directed transepithelial PMN migration ensued. In contrast, attachment of a non-pathogenic Escherichia coli strain to the apical membrane of epithelial cells at comparable densities failed to stimulate a directed PMN transepithelial migration. Use of the n-formyl-peptide receptor antagonist N-t-BOC-1-methionyl-1-leucyl-1- phenylalanine (tBOC-MLP) indicated that the Salmonella-induced PMN transepithelial migration response was not attributable to the classical pathway by which bacteria induce directed migration of PMN. Moreover, the PMN transmigration response required Salmonella adhesion to the epithelial apical membrane and subsequent reciprocal protein synthesis in both bacteria and epithelial cells. Among the events stimulated by this interaction was the epithelial synthesis and polarized release of the potent PMN chemotactic peptide interleukin-8 (IL-8). However, IL-8 neutralization, transfer, and induction experiments indicated that this cytokine was not responsible for the elicited PMN transmigration. These data indicate that a novel transcellular pathway exists in which subepithelial PMN respond to lumenal pathogens across a functionally intact epithelium. Based on the known unique characteristics of the intestinal mucosa, we speculate that IL-8 may act in concert with an as yet unidentified transcellular chemotactic factor(s) (TCF) which directs PMN migration across the intestinal epithelium.

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Selected References

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  1. Agace W., Hedges S., Andersson U., Andersson J., Ceska M., Svanborg C. Selective cytokine production by epithelial cells following exposure to Escherichia coli. Infect Immun. 1993 Feb;61(2):602–609. doi: 10.1128/iai.61.2.602-609.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alpuche Aranda C. M., Swanson J. A., Loomis W. P., Miller S. I. Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10079–10083. doi: 10.1073/pnas.89.21.10079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Butcher E. C. Leukocyte-endothelial cell recognition: three (or more) steps to specificity and diversity. Cell. 1991 Dec 20;67(6):1033–1036. doi: 10.1016/0092-8674(91)90279-8. [DOI] [PubMed] [Google Scholar]
  4. Chadwick V. S., Mellor D. M., Myers D. B., Selden A. C., Keshavarzian A., Broom M. F., Hobson C. H. Production of peptides inducing chemotaxis and lysosomal enzyme release in human neutrophils by intestinal bacteria in vitro and in vivo. Scand J Gastroenterol. 1988 Jan;23(1):121–128. doi: 10.3109/00365528809093861. [DOI] [PubMed] [Google Scholar]
  5. Cohen P. S., Rossoll R., Cabelli V. J., Yang S. L., Laux D. C. Relationship between the mouse colonizing ability of a human fecal Escherichia coli strain and its ability to bind a specific mouse colonic mucous gel protein. Infect Immun. 1983 Apr;40(1):62–69. doi: 10.1128/iai.40.1.62-69.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colgan S. P., Parkos C. A., Delp C., Arnaout M. A., Madara J. L. Neutrophil migration across cultured intestinal epithelial monolayers is modulated by epithelial exposure to IFN-gamma in a highly polarized fashion. J Cell Biol. 1993 Feb;120(3):785–798. doi: 10.1083/jcb.120.3.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cramer E. B., Milks L. C., Brontoli M. J., Ojakian G. K., Wright S. D., Showell H. J. Effect of human serum and some of its components on neutrophil adherence and migration across an epithelium. J Cell Biol. 1986 May;102(5):1868–1877. doi: 10.1083/jcb.102.5.1868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dana N., Todd R. F., 3rd, Pitt J., Springer T. A., Arnaout M. A. Deficiency of a surface membrane glycoprotein (Mo1) in man. J Clin Invest. 1984 Jan;73(1):153–159. doi: 10.1172/JCI111186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Day D. W., Mandal B. K., Morson B. C. The rectal biopsy appearances in Salmonella colitis. Histopathology. 1978 Mar;2(2):117–131. doi: 10.1111/j.1365-2559.1978.tb01700.x. [DOI] [PubMed] [Google Scholar]
  10. Dharmsathaphorn K., Madara J. L. Established intestinal cell lines as model systems for electrolyte transport studies. Methods Enzymol. 1990;192:354–389. doi: 10.1016/0076-6879(90)92082-o. [DOI] [PubMed] [Google Scholar]
  11. Ernst R. K., Dombroski D. M., Merrick J. M. Anaerobiosis, type 1 fimbriae, and growth phase are factors that affect invasion of HEp-2 cells by Salmonella typhimurium. Infect Immun. 1990 Jun;58(6):2014–2016. doi: 10.1128/iai.58.6.2014-2016.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Evans C. W., Taylor J. E., Walker J. D., Simmons N. L. Transepithelial chemotaxis of rat peritoneal exudate cells. Br J Exp Pathol. 1983 Dec;64(6):644–654. [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Finlay B. B., Gumbiner B., Falkow S. Penetration of Salmonella through a polarized Madin-Darby canine kidney epithelial cell monolayer. J Cell Biol. 1988 Jul;107(1):221–230. doi: 10.1083/jcb.107.1.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Galán J. E., Curtiss R., 3rd Expression of Salmonella typhimurium genes required for invasion is regulated by changes in DNA supercoiling. Infect Immun. 1990 Jun;58(6):1879–1885. doi: 10.1128/iai.58.6.1879-1885.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Galán J. E., Ginocchio C., Costeas P. Molecular and functional characterization of the Salmonella invasion gene invA: homology of InvA to members of a new protein family. J Bacteriol. 1992 Jul;174(13):4338–4349. doi: 10.1128/jb.174.13.4338-4349.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Galán J. E., Pace J., Hayman M. J. Involvement of the epidermal growth factor receptor in the invasion of cultured mammalian cells by Salmonella typhimurium. Nature. 1992 Jun 18;357(6379):588–589. doi: 10.1038/357588a0. [DOI] [PubMed] [Google Scholar]
  18. Hedges S., Anderson P., Lidin-Janson G., de Man P., Svanborg C. Interleukin-6 response to deliberate colonization of the human urinary tract with gram-negative bacteria. Infect Immun. 1991 Jan;59(1):421–427. doi: 10.1128/iai.59.1.421-427.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Huber A. R., Kunkel S. L., Todd R. F., 3rd, Weiss S. J. Regulation of transendothelial neutrophil migration by endogenous interleukin-8. Science. 1991 Oct 4;254(5028):99–102. doi: 10.1126/science.1718038. [DOI] [PubMed] [Google Scholar]
  21. Iványi B., Ormos J., Lantos J. Tubulointerstitial inflammation, cast formation, and renal parenchymal damage in experimental pyelonephritis. Am J Pathol. 1983 Dec;113(3):300–308. [PMC free article] [PubMed] [Google Scholar]
  22. Jirik F. R., Podor T. J., Hirano T., Kishimoto T., Loskutoff D. J., Carson D. A., Lotz M. Bacterial lipopolysaccharide and inflammatory mediators augment IL-6 secretion by human endothelial cells. J Immunol. 1989 Jan 1;142(1):144–147. [PubMed] [Google Scholar]
  23. Jodal M., Lundgren O. Countercurrent mechanisms in the mammalian gastrointestinal tract. Gastroenterology. 1986 Jul;91(1):225–241. doi: 10.1016/0016-5085(86)90463-4. [DOI] [PubMed] [Google Scholar]
  24. Ko Y. C., Mukaida N., Ishiyama S., Tokue A., Kawai T., Matsushima K., Kasahara T. Elevated interleukin-8 levels in the urine of patients with urinary tract infections. Infect Immun. 1993 Apr;61(4):1307–1314. doi: 10.1128/iai.61.4.1307-1314.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Koyama S., Rennard S. I., Leikauf G. D., Shoji S., Von Essen S., Claassen L., Robbins R. A. Endotoxin stimulates bronchial epithelial cells to release chemotactic factors for neutrophils. A potential mechanism for neutrophil recruitment, cytotoxicity, and inhibition of proliferation in bronchial inflammation. J Immunol. 1991 Dec 15;147(12):4293–4301. [PubMed] [Google Scholar]
  27. Kumar N. B., Nostrant T. T., Appelman H. D. The histopathologic spectrum of acute self-limited colitis (acute infectious-type colitis). Am J Surg Pathol. 1982 Sep;6(6):523–529. doi: 10.1097/00000478-198209000-00004. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Lissner C. R., Swanson R. N., O'Brien A. D. Genetic control of the innate resistance of mice to Salmonella typhimurium: expression of the Ity gene in peritoneal and splenic macrophages isolated in vitro. J Immunol. 1983 Dec;131(6):3006–3013. [PubMed] [Google Scholar]
  30. Loppnow H., Libby P., Freudenberg M., Krauss J. H., Weckesser J., Mayer H. Cytokine induction by lipopolysaccharide (LPS) corresponds to lethal toxicity and is inhibited by nontoxic Rhodobacter capsulatus LPS. Infect Immun. 1990 Nov;58(11):3743–3750. doi: 10.1128/iai.58.11.3743-3750.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Luscinskas F. W., Brock A. F., Arnaout M. A., Gimbrone M. A., Jr Endothelial-leukocyte adhesion molecule-1-dependent and leukocyte (CD11/CD18)-dependent mechanisms contribute to polymorphonuclear leukocyte adhesion to cytokine-activated human vascular endothelium. J Immunol. 1989 Apr 1;142(7):2257–2263. [PubMed] [Google Scholar]
  32. Madara J. L., Dharmsathaphorn K. Occluding junction structure-function relationships in a cultured epithelial monolayer. J Cell Biol. 1985 Dec;101(6):2124–2133. doi: 10.1083/jcb.101.6.2124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Madara J. L., Parkos C., Colgan S., MacLeod R. J., Nash S., Matthews J., Delp C., Lencer W. Cl- secretion in a model intestinal epithelium induced by a neutrophil-derived secretagogue. J Clin Invest. 1992 Jun;89(6):1938–1944. doi: 10.1172/JCI115800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Madara J. L., Patapoff T. W., Gillece-Castro B., Colgan S. P., Parkos C. A., Delp C., Mrsny R. J. 5'-adenosine monophosphate is the neutrophil-derived paracrine factor that elicits chloride secretion from T84 intestinal epithelial cell monolayers. J Clin Invest. 1993 May;91(5):2320–2325. doi: 10.1172/JCI116462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Madara J. L. Warner-Lambert/Parke-Davis Award lecture. Pathobiology of the intestinal epithelial barrier. Am J Pathol. 1990 Dec;137(6):1273–1281. [PMC free article] [PubMed] [Google Scholar]
  36. McCormick B. A., Franklin D. P., Laux D. C., Cohen P. S. Type 1 pili are not necessary for colonization of the streptomycin-treated mouse large intestine by type 1-piliated Escherichia coli F-18 and E. coli K-12. Infect Immun. 1989 Oct;57(10):3022–3029. doi: 10.1128/iai.57.10.3022-3029.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. McGovern V. J., Slavutin L. J. Pathology of salmonella colitis. Am J Surg Pathol. 1979 Dec;3(6):483–490. doi: 10.1097/00000478-197912000-00001. [DOI] [PubMed] [Google Scholar]
  38. McRoberts J. A., Aranda R., Riley N., Kang H. Insulin regulates the paracellular permeability of cultured intestinal epithelial cell monolayers. J Clin Invest. 1990 Apr;85(4):1127–1134. doi: 10.1172/JCI114544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Migliorisi G., Folkes E., Cramer E. B. Differences in the ability of neutrophils and monocytes to traverse epithelial occluding junctions. J Leukoc Biol. 1988 Dec;44(6):485–492. doi: 10.1002/jlb.44.6.485. [DOI] [PubMed] [Google Scholar]
  40. Myhal M. L., Laux D. C., Cohen P. S. Relative colonizing abilities of human fecal and K 12 strains of Escherichia coli in the large intestines of streptomycin-treated mice. Eur J Clin Microbiol. 1982 Jun;1(3):186–192. doi: 10.1007/BF02019621. [DOI] [PubMed] [Google Scholar]
  41. Nash S., Parkos C., Nusrat A., Delp C., Madara J. L. In vitro model of intestinal crypt abscess. A novel neutrophil-derived secretagogue activity. J Clin Invest. 1991 Apr;87(4):1474–1477. doi: 10.1172/JCI115156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Nash S., Stafford J., Madara J. L. Effects of polymorphonuclear leukocyte transmigration on the barrier function of cultured intestinal epithelial monolayers. J Clin Invest. 1987 Oct;80(4):1104–1113. doi: 10.1172/JCI113167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Parkos C. A., Colgan S. P., Delp C., Arnaout M. A., Madara J. L. Neutrophil migration across a cultured epithelial monolayer elicits a biphasic resistance response representing sequential effects on transcellular and paracellular pathways. J Cell Biol. 1992 May;117(4):757–764. doi: 10.1083/jcb.117.4.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Parkos C. A., Delp C., Arnaout M. A., Madara J. L. Neutrophil migration across a cultured intestinal epithelium. Dependence on a CD11b/CD18-mediated event and enhanced efficiency in physiological direction. J Clin Invest. 1991 Nov;88(5):1605–1612. doi: 10.1172/JCI115473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Parsons P. E., Sugahara K., Cott G. R., Mason R. J., Henson P. M. The effect of neutrophil migration and prolonged neutrophil contact on epithelial permeability. Am J Pathol. 1987 Nov;129(2):302–312. [PMC free article] [PubMed] [Google Scholar]
  46. Pospischil A., Wood R. L., Anderson T. D. Peroxidase-antiperoxidase and immunogold labeling of Salmonella typhimurium and Salmonella choleraesuis var kunzendorf in tissues of experimentally infected swine. Am J Vet Res. 1990 Apr;51(4):619–624. [PubMed] [Google Scholar]
  47. Revel H. R. Restriction of nonglucosylated T-even bacteriophage: properties of permissive mutants of Escherichia coli B and K12. Virology. 1967 Apr;31(4):688–701. doi: 10.1016/0042-6822(67)90197-3. [DOI] [PubMed] [Google Scholar]
  48. Rout W. R., Formal S. B., Dammin G. J., Giannella R. A. Pathophysiology of Salmonella diarrhea in the Rhesus monkey: Intestinal transport, morphological and bacteriological studies. Gastroenterology. 1974 Jul;67(1):59–70. [PubMed] [Google Scholar]
  49. Schiemann D. A., Shope S. R. Anaerobic growth of Salmonella typhimurium results in increased uptake by Henle 407 epithelial and mouse peritoneal cells in vitro and repression of a major outer membrane protein. Infect Immun. 1991 Jan;59(1):437–440. doi: 10.1128/iai.59.1.437-440.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Smith C. W., Rothlein R., Hughes B. J., Mariscalco M. M., Rudloff H. E., Schmalstieg F. C., Anderson D. C. Recognition of an endothelial determinant for CD 18-dependent human neutrophil adherence and transendothelial migration. J Clin Invest. 1988 Nov;82(5):1746–1756. doi: 10.1172/JCI113788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Takada H., Mihara J., Morisaki I., Hamada S. Induction of interleukin-1 and -6 in human gingival fibroblast cultures stimulated with Bacteroides lipopolysaccharides. Infect Immun. 1991 Jan;59(1):295–301. doi: 10.1128/iai.59.1.295-301.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]
  53. Teahon K., Smethurst P., Pearson M., Levi A. J., Bjarnason I. The effect of elemental diet on intestinal permeability and inflammation in Crohn's disease. Gastroenterology. 1991 Jul;101(1):84–89. doi: 10.1016/0016-5085(91)90463-u. [DOI] [PubMed] [Google Scholar]
  54. Tsujimura S., Otaguro K., Sasaki M., Tanaka R. Chemotactic factors as cause of polyleukocyte urine in urinary tract infection: chemotactic activity of urinary protein fraction. Invest Urol. 1980 Jan;17(4):269–272. [PubMed] [Google Scholar]
  55. Wolf J. L., Kauffman R. S., Finberg R., Dambrauskas R., Fields B. N., Trier J. S. Determinants of reovirus interaction with the intestinal M cells and absorptive cells of murine intestine. Gastroenterology. 1983 Aug;85(2):291–300. [PubMed] [Google Scholar]

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