Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Jul;65(7):2950–2958. doi: 10.1128/iai.65.7.2950-2958.1997

Invasion of cultured human epithelial cells by Klebsiella pneumoniae isolated from the urinary tract.

T A Oelschlaeger 1, B D Tall 1
PMCID: PMC175413  PMID: 9199471

Abstract

The mechanisms which enable entry into cultured human epithelial cells by Klebsiella pneumoniae were compared with those of Salmonella typhi Ty2. K. pneumoniae 3091, isolated from a urine sample of a patient with a urinary tract infection, invaded human epithelial cells from the bladder and ileocecum and persisted for days in vitro. Electron microscopic studies demonstrated that K. pneumoniae was always contained in endosomes. The internalization mechanism(s) triggered by K. pneumoniae was studied by invasion assays conducted with different inhibitors that act on prokaryotic and eukaryotic cell structures and processes. Chloramphenicol inhibition of bacterial uptake revealed that bacterial de novo protein synthesis was essential for efficient invasion by K. pneumoniae and S. typhi. Interference with receptor-mediated endocytosis by g-strophanthin or monodansylcadaverine and inhibition of endosome acidification by monensin reduced the number of viable intracellular K. pneumoniae cells, but not S. typhi cells. The depolymerization of microfilaments by cytochalasin D inhibited the uptake of both bacteria. Microtubule depolymerization caused by colchicine, demecolcine, or nocodazole and the stabilization of microtubules with taxol reduced only the invasion ability of K. pneumoniae. S. typhi invasion was unaffected by microtubule depolymerization or stabilization. These data suggest that the internalization mechanism triggered by K. pneumoniae 3091 is strikingly different from the solely microfilament-dependent invasion mechanism exhibited by many of the well-studied enteric bacteria, such as enteroinvasive Escherichia coli, Salmonella, Shigella, and Yersinia strains.

Full Text

The Full Text of this article is available as a PDF (656.6 KB).

Selected References

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

  1. Arlet G., Rouveau M., Casin I., Bouvet P. J., Lagrange P. H., Philippon A. Molecular epidemiology of Klebsiella pneumoniae strains that produce SHV-4 beta-lactamase and which were isolated in 14 French hospitals. J Clin Microbiol. 1994 Oct;32(10):2553–2558. doi: 10.1128/jcm.32.10.2553-2558.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Basu S. K., Goldstein J. L., Anderson R. G., Brown M. S. Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts. Cell. 1981 May;24(2):493–502. doi: 10.1016/0092-8674(81)90340-8. [DOI] [PubMed] [Google Scholar]
  3. Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Courtney M. A., Miller J. R., Summersgill J., Melo J., Raff M. J., Streips U. N. R-factor responsible for an outbreak of multiply antibiotic-resistant Klebsiella pneumoniae. Antimicrob Agents Chemother. 1980 Dec;18(6):926–929. doi: 10.1128/aac.18.6.926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DUGUID J. P. Fimbriae and adhesive properties in Klebsiella strains. J Gen Microbiol. 1959 Aug;21:271–286. doi: 10.1099/00221287-21-1-271. [DOI] [PubMed] [Google Scholar]
  6. Darfeuille-Michaud A., Jallat C., Aubel D., Sirot D., Rich C., Sirot J., Joly B. R-plasmid-encoded adhesive factor in Klebsiella pneumoniae strains responsible for human nosocomial infections. Infect Immun. 1992 Jan;60(1):44–55. doi: 10.1128/iai.60.1.44-55.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Davies P. J., Davies D. R., Levitzki A., Maxfield F. R., Milhaud P., Willingham M. C., Pastan I. H. Transglutaminase is essential in receptor-mediated endocytosis of alpha 2-macroglobulin and polypeptide hormones. Nature. 1980 Jan 10;283(5743):162–167. doi: 10.1038/283162a0. [DOI] [PubMed] [Google Scholar]
  8. De Brabander M., Geuens G., Nuydens R., Willebrords R., De Mey J. Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5608–5612. doi: 10.1073/pnas.78.9.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dean R. T., Jessup W., Roberts C. R. Effects of exogenous amines on mammalian cells, with particular reference to membrane flow. Biochem J. 1984 Jan 1;217(1):27–40. doi: 10.1042/bj2170027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Detilleux P. G., Deyoe B. L., Cheville N. F. Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells. Am J Vet Res. 1991 Oct;52(10):1658–1664. [PubMed] [Google Scholar]
  11. Donnenberg M. S., Donohue-Rolfe A., Keusch G. T. A comparison of HEp-2 cell invasion by enteropathogenic and enteroinvasive Escherichia coli. FEMS Microbiol Lett. 1990 May;57(1-2):83–86. doi: 10.1016/0378-1097(90)90417-o. [DOI] [PubMed] [Google Scholar]
  12. Elsinghorst E. A., Baron L. S., Kopecko D. J. Penetration of human intestinal epithelial cells by Salmonella: molecular cloning and expression of Salmonella typhi invasion determinants in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5173–5177. doi: 10.1073/pnas.86.13.5173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evans D. G., Evans D. J., Jr, Graham D. Y. Adherence and internalization of Helicobacter pylori by HEp-2 cells. Gastroenterology. 1992 May;102(5):1557–1567. doi: 10.1016/0016-5085(92)91714-f. [DOI] [PubMed] [Google Scholar]
  14. Fader R. C., Avots-Avotins A. E., Davis C. P. Evidence for pili-mediated adherence of Klebsiella pneumoniae to rat bladder epithelial cells in vitro. Infect Immun. 1979 Aug;25(2):729–737. doi: 10.1128/iai.25.2.729-737.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fader R. C., Gondesen K., Tolley B., Ritchie D. G., Moller P. Evidence that in vitro adherence of Klebsiella pneumoniae to ciliated hamster tracheal cells is mediated by type 1 fimbriae. Infect Immun. 1988 Nov;56(11):3011–3013. doi: 10.1128/iai.56.11.3011-3013.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Finlay B. B., Falkow S. Common themes in microbial pathogenicity. Microbiol Rev. 1989 Jun;53(2):210–230. doi: 10.1128/mr.53.2.210-230.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Finlay B. B., Falkow S. Comparison of the invasion strategies used by Salmonella cholerae-suis, Shigella flexneri and Yersinia enterocolitica to enter cultured animal cells: endosome acidification is not required for bacterial invasion or intracellular replication. Biochimie. 1988 Aug;70(8):1089–1099. doi: 10.1016/0300-9084(88)90271-4. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Grüneberg R. N. Changes in the antibiotic sensitivities of urinary pathogens, 1971-1989. J Antimicrob Chemother. 1990 Dec;26 (Suppl F):3–11. doi: 10.1093/jac/26.suppl_f.3. [DOI] [PubMed] [Google Scholar]
  20. Hodinka R. L., Wyrick P. B. Ultrastructural study of mode of entry of Chlamydia psittaci into L-929 cells. Infect Immun. 1986 Dec;54(3):855–863. doi: 10.1128/iai.54.3.855-863.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Larkin J. M., Brown M. S., Goldstein J. L., Anderson R. G. Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts. Cell. 1983 May;33(1):273–285. doi: 10.1016/0092-8674(83)90356-2. [DOI] [PubMed] [Google Scholar]
  22. Meier C., Oelschlaeger T. A., Merkert H., Korhonen T. K., Hacker J. Ability of Escherichia coli isolates that cause meningitis in newborns to invade epithelial and endothelial cells. Infect Immun. 1996 Jul;64(7):2391–2399. doi: 10.1128/iai.64.7.2391-2399.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Merino S., Camprubí S., Albertí S., Benedí V. J., Tomás J. M. Mechanisms of Klebsiella pneumoniae resistance to complement-mediated killing. Infect Immun. 1992 Jun;60(6):2529–2535. doi: 10.1128/iai.60.6.2529-2535.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miliotis M. D. Acridine orange stain for determining intracellular enteropathogens in HeLa cells. J Clin Microbiol. 1991 Apr;29(4):830–831. doi: 10.1128/jcm.29.4.830-831.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moya M., Dautry-Varsat A., Goud B., Louvard D., Boquet P. Inhibition of coated pit formation in Hep2 cells blocks the cytotoxicity of diphtheria toxin but not that of ricin toxin. J Cell Biol. 1985 Aug;101(2):548–559. doi: 10.1083/jcb.101.2.548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nassif X., Sansonetti P. J. Correlation of the virulence of Klebsiella pneumoniae K1 and K2 with the presence of a plasmid encoding aerobactin. Infect Immun. 1986 Dec;54(3):603–608. doi: 10.1128/iai.54.3.603-608.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oelschlaeger T. A., Barrett T. J., Kopecko D. J. Some structures and processes of human epithelial cells involved in uptake of enterohemorrhagic Escherichia coli O157:H7 strains. Infect Immun. 1994 Nov;62(11):5142–5150. doi: 10.1128/iai.62.11.5142-5150.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Oelschlaeger T. A., Guerry P., Kopecko D. J. Unusual microtubule-dependent endocytosis mechanisms triggered by Campylobacter jejuni and Citrobacter freundii. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6884–6888. doi: 10.1073/pnas.90.14.6884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oelschlaeger T. A., Tall B. D. Uptake pathways of clinical isolates of Proteus mirabilis into human epithelial cell lines. Microb Pathog. 1996 Jul;21(1):1–16. doi: 10.1006/mpat.1996.0037. [DOI] [PubMed] [Google Scholar]
  30. Old D. C., Adegbola R. A. A new mannose-resistant haemagglutinin in Klebsiella. J Appl Bacteriol. 1983 Aug;55(1):165–172. doi: 10.1111/j.1365-2672.1983.tb02661.x. [DOI] [PubMed] [Google Scholar]
  31. Old D. C. Inhibition of the interaction between fimbrial haemagglutinins and erythrocytes by D-mannose and other carbohydrates. J Gen Microbiol. 1972 Jun;71(1):149–157. doi: 10.1099/00221287-71-1-149. [DOI] [PubMed] [Google Scholar]
  32. Osborn M., Weber K. Immunofluorescence and immunocytochemical procedures with affinity purified antibodies: tubulin-containing structures. Methods Cell Biol. 1982;24:97–132. doi: 10.1016/s0091-679x(08)60650-0. [DOI] [PubMed] [Google Scholar]
  33. Pruzzo C., Debbia E. A., Satta G. Identification of the major adherence ligand of Klebsiella pneumoniae in the receptor for coliphage T7 and alteration of Klebsiella adherence properties by lysogenic conversion. Infect Immun. 1980 Nov;30(2):562–571. doi: 10.1128/iai.30.2.562-571.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Przondo Hessek A., Pulverer G. Hemagglutinins of Klebsiella pneumoniae and Klebsiella oxytoca. Zentralbl Bakteriol Mikrobiol Hyg A. 1983 Nov;255(4):472–478. [PubMed] [Google Scholar]
  35. Reynolds D. J., Pearce J. H. Characterization of the cytochalasin D-resistant (pinocytic) mechanisms of endocytosis utilized by chlamydiae. Infect Immun. 1990 Oct;58(10):3208–3216. doi: 10.1128/iai.58.10.3208-3216.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Richardson W. P., Sadoff J. C. Induced engulfment of Neisseria gonorrhoeae by tissue culture cells. Infect Immun. 1988 Sep;56(9):2512–2514. doi: 10.1128/iai.56.9.2512-2514.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schiff P. B., Horwitz S. B. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1561–1565. doi: 10.1073/pnas.77.3.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schwartz A. L., Bolognesi A., Fridovich S. E. Recycling of the asialoglycoprotein receptor and the effect of lysosomotropic amines in hepatoma cells. J Cell Biol. 1984 Feb;98(2):732–738. doi: 10.1083/jcb.98.2.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Selden R., Lee S., Wang W. L., Bennett J. V., Eickhoff T. C. Nosocomial klebsiella infections: intestinal colonization as a reservoir. Ann Intern Med. 1971 May;74(5):657–664. doi: 10.7326/0003-4819-74-5-657. [DOI] [PubMed] [Google Scholar]
  40. St Geme J. W., 3rd, Falkow S. Haemophilus influenzae adheres to and enters cultured human epithelial cells. Infect Immun. 1990 Dec;58(12):4036–4044. doi: 10.1128/iai.58.12.4036-4044.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stamm W. E., McKevitt M., Roberts P. L., White N. J. Natural history of recurrent urinary tract infections in women. Rev Infect Dis. 1991 Jan-Feb;13(1):77–84. doi: 10.1093/clinids/13.1.77. [DOI] [PubMed] [Google Scholar]
  42. Straus D. C. Production of an extracellular toxic complex by various strains of Klebsiella pneumoniae. Infect Immun. 1987 Jan;55(1):44–48. doi: 10.1128/iai.55.1.44-48.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Söderlund G., Kihlström E. Effect of methylamine and monodansylcadaverine on the susceptibility of McCoy cells to Chlamydia trachomatis infection. Infect Immun. 1983 May;40(2):534–541. doi: 10.1128/iai.40.2.534-541.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Van Leuven F., Cassiman J. J., Van Den Berghe H. Primary amines inhibit recycling of alpha 2M receptors in fibroblasts. Cell. 1980 May;20(1):37–43. doi: 10.1016/0092-8674(80)90232-9. [DOI] [PubMed] [Google Scholar]
  45. Ward M. E., Murray A. Control mechanisms governing the infectivity of Chlamydia trachomatis for HeLa cells: mechanisms of endocytosis. J Gen Microbiol. 1984 Jul;130(7):1765–1780. doi: 10.1099/00221287-130-7-1765. [DOI] [PubMed] [Google Scholar]
  46. Wyrick P. B., Choong J., Davis C. H., Knight S. T., Royal M. O., Maslow A. S., Bagnell C. R. Entry of genital Chlamydia trachomatis into polarized human epithelial cells. Infect Immun. 1989 Aug;57(8):2378–2389. doi: 10.1128/iai.57.8.2378-2389.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES