Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1994 Oct 1;180(4):1437–1443. doi: 10.1084/jem.180.4.1437

Endotoxin "priming" potentiates lung vascular abnormalities in response to Escherichia coli hemolysin: an example of synergism between endo- and exotoxin

PMCID: PMC2191678  PMID: 7931076

Abstract

The pore-forming hemolysin of Escherichia coli (HlyA), an important virulence factor in extraintestinal E. coli infections, causes thromboxane generation and related vasoconstriction in perfused rabbit lungs (Seeger, W., H. Walter, N. Suttorp, M. Muhly, and S. Bhakdi. 1989. J. Clin. Invest. 84:220). We investigated the influence of pulmonary vascular "priming" with endotoxin on the responsiveness of the lung to a low-dose HlyA challenge. Rabbit lungs were perfused with Krebs Henseleit buffer containing 0.1-100 ng/ml Salmonella abortus equii lipopolysaccharide (LPS) for 60-180 min. This treatment caused protracted release of tumor necrosis factor into the recirculating medium, but did not induce significant alterations of pulmonary hemodynamics and fluid balance. At a dose of 1 ng/ml, HlyA elicited only moderate thromboxane release (< 200 pg/ml) and pulmonary artery pressure increase (< or = 6 mmHg) in control lungs. Acceleration and potentiation of both the metabolic and vasoconstrictor response occurred in lungs primed with LPS. This priming effect displayed dose (threshold integral of 0.1-1 ng/ml LPS) and time dependencies (threshold integral of 60-90 min LPS incubation). Maximum thromboxane release and pulmonary artery pressure increase surpassed the responses to HlyA in nonprimed lungs by more than 15-fold. Cyclooxygenase inhibition and thromboxane-receptor antagonism blocked these effects. These data demonstrate that LPS priming synergizes with HlyA challenge to provoke vascular abnormalities that are possibly relevant to the pathogenesis of organ failure in severe local and systemic infections.

Full Text

The Full Text of this article is available as a PDF (649.9 KB).

Selected References

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

  1. Aderem A. A., Cohen D. S., Wright S. D., Cohn Z. A. Bacterial lipopolysaccharides prime macrophages for enhanced release of arachidonic acid metabolites. J Exp Med. 1986 Jul 1;164(1):165–179. doi: 10.1084/jem.164.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aderem A. A., Cohn Z. A. Calcium ionophore synergizes with bacterial lipopolysaccharides in activating macrophage arachidonic acid metabolism. J Exp Med. 1988 Feb 1;167(2):623–631. doi: 10.1084/jem.167.2.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhakdi S., Martin E. Superoxide generation by human neutrophils induced by low doses of Escherichia coli hemolysin. Infect Immun. 1991 Sep;59(9):2955–2962. doi: 10.1128/iai.59.9.2955-2962.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhakdi S., Muhly M., Korom S., Schmidt G. Effects of Escherichia coli hemolysin on human monocytes. Cytocidal action and stimulation of interleukin 1 release. J Clin Invest. 1990 Jun;85(6):1746–1753. doi: 10.1172/JCI114631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bhakdi S., Muhly M., Mannhardt U., Hugo F., Klapettek K., Mueller-Eckhardt C., Roka L. Staphylococcal alpha toxin promotes blood coagulation via attack on human platelets. J Exp Med. 1988 Aug 1;168(2):527–542. doi: 10.1084/jem.168.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bhakdi S., Weller U., Walev I., Martin E., Jonas D., Palmer M. A guide to the use of pore-forming toxins for controlled permeabilization of cell membranes. Med Microbiol Immunol. 1993 Sep;182(4):167–175. doi: 10.1007/BF00219946. [DOI] [PubMed] [Google Scholar]
  7. Braun V., Focareta T. Pore-forming bacterial protein hemolysins (cytolysins). Crit Rev Microbiol. 1991;18(2):115–158. doi: 10.3109/10408419109113511. [DOI] [PubMed] [Google Scholar]
  8. Brigham K. L., Meyrick B. Endotoxin and lung injury. Am Rev Respir Dis. 1986 May;133(5):913–927. [PubMed] [Google Scholar]
  9. Brigham K. L., Meyrick B. Endotoxin and lung injury. Am Rev Respir Dis. 1986 May;133(5):913–927. [PubMed] [Google Scholar]
  10. Cavalieri S. J., Bohach G. A., Snyder I. S. Escherichia coli alpha-hemolysin: characteristics and probable role in pathogenicity. Microbiol Rev. 1984 Dec;48(4):326–343. doi: 10.1128/mr.48.4.326-343.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Christman J. W., Petras S. F., Vacek P. M., Davis G. S. Rat alveolar macrophage production of chemoattractants for neutrophils: response to Escherichia coli endotoxin. Infect Immun. 1989 Mar;57(3):810–816. doi: 10.1128/iai.57.3.810-816.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Doerfler M. E., Danner R. L., Shelhamer J. H., Parrillo J. E. Bacterial lipopolysaccharides prime human neutrophils for enhanced production of leukotriene B4. J Clin Invest. 1989 Mar;83(3):970–977. doi: 10.1172/JCI113983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eberspächer B., Hugo F., Pohl M., Bhakdi S. Functional similarity between the haemolysins of Escherichia coli and Morganella morganii. J Med Microbiol. 1990 Nov;33(3):165–170. doi: 10.1099/00222615-33-3-165. [DOI] [PubMed] [Google Scholar]
  14. Ermert L., Seeger W., Duncker H. R. Computer-assisted morphometry of the intracapillary leukocyte pool in the rabbit lung. Cell Tissue Res. 1993 Mar;271(3):469–476. doi: 10.1007/BF02913729. [DOI] [PubMed] [Google Scholar]
  15. Feuerstein N., Ramwell P. W. In vivo and in vitro effects of endotoxin on prostaglandin release from rat lung. Br J Pharmacol. 1981 Jun;73(2):511–516. doi: 10.1111/j.1476-5381.1981.tb10450.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Forehand J. R., Pabst M. J., Phillips W. A., Johnston R. B., Jr Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium. J Clin Invest. 1989 Jan;83(1):74–83. doi: 10.1172/JCI113887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fünfstück R., Tschäpe H., Stein G., Kunath H., Bergner M., Wessel G. Virulence properties of Escherichia coli strains in patients with chronic pyelonephritis. Infection. 1986 May-Jun;14(3):145–150. doi: 10.1007/BF01643482. [DOI] [PubMed] [Google Scholar]
  18. Goldman D. W., Enkel H., Gifford L. A., Chenoweth D. E., Rosenbaum J. T. Lipopolysaccharide modulates receptors for leukotriene B4, C5a, and formyl-methionyl-leucyl-phenylalanine on rabbit polymorphonuclear leukocytes. J Immunol. 1986 Sep 15;137(6):1971–1976. [PubMed] [Google Scholar]
  19. Grimminger F., Becker G., Seeger W. High yield enzymatic conversion of intravascular leukotriene A4 in blood-free perfused lungs. J Immunol. 1988 Oct 1;141(7):2431–2436. [PubMed] [Google Scholar]
  20. Grimminger F., Scholz C., Bhakdi S., Seeger W. Subhemolytic doses of Escherichia coli hemolysin evoke large quantities of lipoxygenase products in human neutrophils. J Biol Chem. 1991 Aug 5;266(22):14262–14269. [PubMed] [Google Scholar]
  21. Grimminger F., Sibelius U., Bhakdi S., Suttorp N., Seeger W. Escherichia coli hemolysin is a potent inductor of phosphoinositide hydrolysis and related metabolic responses in human neutrophils. J Clin Invest. 1991 Nov;88(5):1531–1539. doi: 10.1172/JCI115463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hacker J., Hughes C., Hof H., Goebel W. Cloned hemolysin genes from Escherichia coli that cause urinary tract infection determine different levels of toxicity in mice. Infect Immun. 1983 Oct;42(1):57–63. doi: 10.1128/iai.42.1.57-63.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Iwase M., Lally E. T., Berthold P., Korchak H. M., Taichman N. S. Effects of cations and osmotic protectants on cytolytic activity of Actinobacillus actinomycetemcomitans leukotoxin. Infect Immun. 1990 Jun;58(6):1782–1788. doi: 10.1128/iai.58.6.1782-1788.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kelley J. Cytokines of the lung. Am Rev Respir Dis. 1990 Mar;141(3):765–788. doi: 10.1164/ajrccm/141.3.765. [DOI] [PubMed] [Google Scholar]
  25. Lo R. Y., Strathdee C. A., Shewen P. E. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1. Infect Immun. 1987 Sep;55(9):1987–1996. doi: 10.1128/iai.55.9.1987-1996.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Magnuson D. K., Maier R. V., Pohlman T. H. Protein kinase C: a potential pathway of endothelial cell activation by endotoxin, tumor necrosis factor, and interleukin-1. Surgery. 1989 Aug;106(2):216–223. [PubMed] [Google Scholar]
  27. Mathison J. C., Wolfson E., Ulevitch R. J. Participation of tumor necrosis factor in the mediation of gram negative bacterial lipopolysaccharide-induced injury in rabbits. J Clin Invest. 1988 Jun;81(6):1925–1937. doi: 10.1172/JCI113540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Olson N. C., Salzer W. L., McCall C. E. Biochemical, physiological and clinical aspects of endotoxemia. Mol Aspects Med. 1988;10(6):511–629. doi: 10.1016/0098-2997(88)90024-6. [DOI] [PubMed] [Google Scholar]
  29. Pabst M. J., Johnston R. B., Jr Increased production of superoxide anion by macrophages exposed in vitro to muramyl dipeptide or lipopolysaccharide. J Exp Med. 1980 Jan 1;151(1):101–114. doi: 10.1084/jem.151.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prpic V., Weiel J. E., Somers S. D., DiGuiseppi J., Gonias S. L., Pizzo S. V., Hamilton T. A., Herman B., Adams D. O. Effects of bacterial lipopolysaccharide on the hydrolysis of phosphatidylinositol-4,5-bisphosphate in murine peritoneal macrophages. J Immunol. 1987 Jul 15;139(2):526–533. [PubMed] [Google Scholar]
  31. Raetz C. R., Ulevitch R. J., Wright S. D., Sibley C. H., Ding A., Nathan C. F. Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J. 1991 Sep;5(12):2652–2660. doi: 10.1096/fasebj.5.12.1916089. [DOI] [PubMed] [Google Scholar]
  32. Repine J. E., Fox R. B., Berger E. M., Vatter A., Shasby D. M., Bowman C. M., Harada R. N. Potential mechanisms of lung injury from hydroxyl radical. Chest. 1981 Jul;80(1 Suppl):45–48. doi: 10.1378/chest.80.1_supplement.45s-a. [DOI] [PubMed] [Google Scholar]
  33. Romano M., Hawiger J. Interaction of endotoxic lipid A and lipid X with purified human platelet protein kinase C. J Biol Chem. 1990 Jan 25;265(3):1765–1770. [PubMed] [Google Scholar]
  34. Salzer W. L., McCall C. E. Primed stimulation of isolated perfused rabbit lung by endotoxin and platelet activating factor induces enhanced production of thromboxane and lung injury. J Clin Invest. 1990 Apr;85(4):1135–1143. doi: 10.1172/JCI114545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Seeger W., Ernst C., Walmrath D., Neuhof H., Roka L. Influence of the thromboxane antagonist BM 13.177 on the arachidonic acid-induced increase in pulmonary vascular resistance and permeability in rabbit lungs. Thromb Res. 1985 Dec 15;40(6):793–805. doi: 10.1016/0049-3848(85)90316-0. [DOI] [PubMed] [Google Scholar]
  36. Seeger W., Obernitz R., Thomas M., Walmrath D., Suttorn N., Holland I. B., Grimminger F., Eberspächer B., Hugo F., Bhakdi S. Lung vascular injury after administration of viable hemolysin-forming Escherichia coli in isolated rabbit lungs. Am Rev Respir Dis. 1991 Apr;143(4 Pt 1):797–805. doi: 10.1164/ajrccm/143.4_Pt_1.797. [DOI] [PubMed] [Google Scholar]
  37. Seeger W., Suttorp N., Schmidt F., Neuhof H. The glutathione redox cycle as a defense system against hydrogen-peroxide-induced prostanoid formation and vasoconstriction in rabbit lungs. Am Rev Respir Dis. 1986 Jun;133(6):1029–1036. doi: 10.1164/arrd.1986.133.6.1029. [DOI] [PubMed] [Google Scholar]
  38. Seeger W., Walmrath D., Grimminger F., Rosseau S., Schütte H., Krämer H. J., Ermert L., Kiss L. Adult respiratory distress syndrome: model systems using isolated perfused rabbit lungs. Methods Enzymol. 1994;233:549–584. doi: 10.1016/s0076-6879(94)33060-3. [DOI] [PubMed] [Google Scholar]
  39. Seeger W., Walmrath D., Menger M., Neuhof H. Increased lung vascular permeability after arachidonic acid and hydrostatic challenge. J Appl Physiol (1985) 1986 Nov;61(5):1781–1789. doi: 10.1152/jappl.1986.61.5.1781. [DOI] [PubMed] [Google Scholar]
  40. Seeger W., Walter H., Suttorp N., Muhly M., Bhakdi S. Thromboxane-mediated hypertension and vascular leakage evoked by low doses of Escherichia coli hemolysin in rabbit lungs. J Clin Invest. 1989 Jul;84(1):220–227. doi: 10.1172/JCI114144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Strathdee C. A., Lo R. Y. Extensive homology between the leukotoxin of Pasteurella haemolytica A1 and the alpha-hemolysin of Escherichia coli. Infect Immun. 1987 Dec;55(12):3233–3236. doi: 10.1128/iai.55.12.3233-3236.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sutherland I. W. Biosynthesis and composition of gram-negative bacterial extracellular and wall polysaccharides. Annu Rev Microbiol. 1985;39:243–270. doi: 10.1146/annurev.mi.39.100185.001331. [DOI] [PubMed] [Google Scholar]
  43. Suttorp N., Flöer B., Schnittler H., Seeger W., Bhakdi S. Effects of Escherichia coli hemolysin on endothelial cell function. Infect Immun. 1990 Nov;58(11):3796–3801. doi: 10.1128/iai.58.11.3796-3801.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Suttorp N., Fuhrmann M., Tannert-Otto S., Grimminger F., Bhadki S. Pore-forming bacterial toxins potently induce release of nitric oxide in porcine endothelial cells. J Exp Med. 1993 Jul 1;178(1):337–341. doi: 10.1084/jem.178.1.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Suttorp N., Galanos C., Neuhof H. Endotoxin alters arachidonate metabolism in pulmonary endothelial cells. Am J Physiol. 1987 Sep;253(3 Pt 1):C384–C390. doi: 10.1152/ajpcell.1987.253.3.C384. [DOI] [PubMed] [Google Scholar]
  46. Szefler S. J., Norton C. E., Ball B., Gross J. M., Aida Y., Pabst M. J. IFN-gamma and LPS overcome glucocorticoid inhibition of priming for superoxide release in human monocytes. Evidence that secretion of IL-1 and tumor necrosis factor-alpha is not essential for monocyte priming. J Immunol. 1989 Jun 1;142(11):3985–3992. [PubMed] [Google Scholar]
  47. Thelen M., Rosen A., Nairn A. C., Aderem A. Tumor necrosis factor alpha modifies agonist-dependent responses in human neutrophils by inducing the synthesis and myristoylation of a specific protein kinase C substrate. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5603–5607. doi: 10.1073/pnas.87.15.5603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Welch R. A., Dellinger E. P., Minshew B., Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. doi: 10.1038/294665a0. [DOI] [PubMed] [Google Scholar]
  49. Welch R. A., Falkow S. Characterization of Escherichia coli hemolysins conferring quantitative differences in virulence. Infect Immun. 1984 Jan;43(1):156–160. doi: 10.1128/iai.43.1.156-160.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Welch R. A. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol. 1991 Mar;5(3):521–528. doi: 10.1111/j.1365-2958.1991.tb00723.x. [DOI] [PubMed] [Google Scholar]
  51. Winn R., Nickelson S., Rice C. L. Fluid filtration coefficient of isolated goat lungs was unchanged by endotoxin. J Appl Physiol (1985) 1988 Jun;64(6):2463–2467. doi: 10.1152/jappl.1988.64.6.2463. [DOI] [PubMed] [Google Scholar]
  52. Wright S. D., Ramos R. A., Tobias P. S., Ulevitch R. J., Mathison J. C. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 1990 Sep 21;249(4975):1431–1433. doi: 10.1126/science.1698311. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES