Modulation of Leukocytes by Exotoxins Produced by HAP Organisms

  • C. J. Czuprynski


The HAP group of organisms is comprised of small Gram-negative rods that can colonize the mucosal surface of the respiratory and genitourinary tracts. In some instances there is a clear relationship between a particular HAP species and its host of choice. In other cases the organism is more promiscuous and will colonize a number of host species. Depending on the HAP species involved, this colonization may be benign, resulting in little or no recognizable infection or host response in a healthy individual (Nicolet, 1990). Subsequent exposure to various types of stressors, or underlying infection with a virus or other pathogenic agent, can favor multiplication of HAP organisms that leads to clinical disease. Leukocytes provide a first line of cellular defense against invading microorganisms. As such, it is critically important for HAP organism to evade the antimicrobial activity of leukocytes if they are to multiply and cause clinical disease. HAP organisms have developed several strategies for doing this, including the production of antiphagocytic capsules and the release of exotoxins (leukotoxins) that inhibit leukocyte antimicrobial activity or kill the leukocytes (Czuprynski and Sample, 1990). More recently, it has become clear that smaller amounts of some of these leukotoxins can activate leukocytes to release oxygen radicals and other inflammatory mediators that likely contribute to the intense inflammation that characterizes many HAP infections (Czuprynski et al., 1991; Breider et al., 1991; Maheswaran et al., 1993; Udeze and Kadis, 1992). The purpose of this chapter is to provide a brief overview of these exotoxins, their relatedness among various HAP species, their differential effects on leukocyte populations, and the possible contributions of other bacterial components (i.e., lipopolysaccharide) to their biological effects.


Alveolar Macrophage Mononuclear Phagocyte Bovine Endothelial Cell Cytocidal Effect Actinobacillus Actinomycetemcomitans 
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  1. Adusu. T.E., Conlon, P.E., Shewen, P.E., and Black, W.D., 1994, Pasteurella haemolvtica leukotoxin induces histamine release from bovine pulmonary mast cells, Can. J. Vet. Res. 58: 1–5.Google Scholar
  2. Bailey, M.J.A., Koronakis, V., Schmoll, T., and Hughes, C., 1992, Escherichia coli HIy T protein, a transcriptional activator of haemolysin synthesis and secretion, is encoded by the ifaH (sfiB) locus required for expression of sex factor and lipopolysaccharide genes. Molec. Microbiol. 6: 1003–1012.Google Scholar
  3. Baluyut, C.S., Simonson, R.R., Bemrick. W.J., and Maheswaran, S.K., 1981, Interaction of Pasteurella haemolvtica with bovine neutrophils: identification and partial characterization of a cytotoxin, Am J. Vet. Res. 42: 1920–1926.Google Scholar
  4. Bauer, M., and Welch, R.A., 1994, Is lipopolysaccharide directly involved in the expression or activity of Escherichia Coli hemolysin? 94th Annu. Meet. Amer. Soc. Microbiol., Abst. B110.Google Scholar
  5. Benson, M.L., Thomson, R.G., and Valli. V.E.O., 1978. The bovine alveolar macrophage. II. In vitro studies with Pasteurella haemolvtica, Can.1. Comp. Med. 42: 368–369.Google Scholar
  6. Berggren, K.A., Baluyut, C.S., Simonson. R.R„ Bemrick. W.J., and Maheswaran, S.K., 1981, Cytotoxic effects of Pasteurella haemolvtica on bovine neutrophils, Am. J. Vet. Res. 42: 1383–1388.Google Scholar
  7. Bhakdi, S. Greulich. S., Muhly, M., Eberspacher, B., Becker, H., Thiele, A., and Hugo, F. 1989, Potent leukocidal action of Escherichia coli hemolysin mediated by permeahilization of target cell membranes, J. Exp. Med. 169: 737–754.PubMedCrossRefGoogle Scholar
  8. Bhakdi, S., Muhly, M. Korom, S., and Schmidt. G. 1990, Effects of Escherichia coli hemolysin on human monocytes, J. Clin. Invest. 85: 1746–1753.Google Scholar
  9. Boehm, D.F., Welch. R.A., and Snyder. I.S., 1990, Domains of Escherichia coli hemolysin (HlyA) involved in binding of calcium and erythrocyte membranes. Infect. Immun. 58: 1959–1964.Google Scholar
  10. Breider, M.A., Kumar. S., and Corstvet, R.E., 1991, interaction of bovine neutrophils in Pasteurella haemolvtica mediated damage to pulmonary endothelial cells, Vet. Immunol. Inmunopathol. 27:337350.Google Scholar
  11. Burrows, L.L., and Lo, R.Y.C., 1992. Molecular characterization of an RTX toxin determinant from Actinobacilius suis. Infect. Immun. 60: 2166–2173.Google Scholar
  12. Byrd, W., and Kadis, S., 1992, Preparation, Characterization, and immunogenicity of conjugate vaccines directed against Actinobacillus pleuropneumoniae virulence determinants, Infect. Immun. 60: 30423051.Google Scholar
  13. Car. B.D., Slauson, D.O., Suyemoto, M.M., Dore, M., and Neilsen, N.R., 1991, Expression and kinetics of induced procoagulant activity in bovine pulmonary alveolar macrophages. Exp. Lung Res. 17: 939957.Google Scholar
  14. Cavalieri, S.J.; and Snyder, I.S., 1982, Effect of Escherichia coli alpha-hemolysin on human peripheral leukocyte function in vitro, Infect. Immun. 37: 966–974.PubMedGoogle Scholar
  15. Chang, Y., Renshaw, H.F., and Richards. A.B., 1986. Pasteurella haemolytica leukotoxin: Physicochemical characteristics and susceptibility of leukotoxin to enzymatic treatment, Am. J. Vet. Res. 47: 716–723.Google Scholar
  16. Chang, Y., Young, R., Post, D., and Struck, K.K., 1987, identification and characteristics of the Pasteurella haemolytica leukotoxin. infect Immun. 55: 2348–2354.Google Scholar
  17. Chung, W.-B. Backstrom. L. McDonald, J., and Collins, M.T„ 1993, Actinobacillus pleuropneumoniae culture supernatants interfere with killing of Pasteurella multocida by swine pulmonary alveolar macrophages, Can. J. Vet. Res. 57:190- I 97.Google Scholar
  18. Clark, R.A., Leidal, K.G., and Taichman, N.S., 1986. Oxidative inactivation of Actinohacillus actinomycetemcomitans leukotoxin by the neutrophil myeloperoxidase system, Infect. Immun. 53: 25 2256.Google Scholar
  19. Clinkenbeard, K.D., Mosier, D.A., and Confer. A.W., 1989, Effects of Pasteurella haemolytica leukotoxin on isolated bovine neutrophils, To. icon 27: 797–804.Google Scholar
  20. Clinkenbeard, K.D., Mosier, D.A., and Confer, A.W., 1989. Transmembrane pore size and role of cell swelling in cytotoxicity caused by Pasteurella haemolytica leukotoxin, infect. Immun. 57: 420–425.PubMedGoogle Scholar
  21. Clinkenbeard, K.D., and Upton, M,L., 1991, Lysis of bovine platelets by Pasteurella haemolytica leukotoxin, Am. J. Vet. Res. 52: 453–457.PubMedGoogle Scholar
  22. Confer, A.W„ Panciera, R.J., Clinkenbeard, K.D., and Mosier, D.A., 1990, Molecular aspects of virulence of Pasteurella haemolytica, Can.J. Vet. Res. 54: S48 - S52.Google Scholar
  23. Cruijsen, T.L.M., Van Leengoed, L.A.M.G., Dekker-Nooren, T.C.E.M., Schoevers, E.J., and Venceijden, J.H.M., 1992, Phagocytosis and killing of Actinohacillus pleuropneumoniae by alveolar macrophages and polymorphonuclear leukocytes isolated from pigs, Infect. Immun. 60: 4867–4871.PubMedGoogle Scholar
  24. Czuprynski, C.J., and Sample, A.K., 1990. Interactions of Ilaemophilus-Actinohacillus-Pasteurella bacteria with phagocytic cells, Can. J. Vet. Res. 54: 536 - S40.Google Scholar
  25. Czuprynski, C.J., Noel, E.J., Ortiz-Carranza, O., and Srikumaran, S., 1991, Activation of bovine neutrophils by partially purified Pasteurella haemolytica leukotoxin, Infect. Immun. 59: 3126–3133.PubMedGoogle Scholar
  26. Czuprynski. C.J., and Ortiz-Carranza. O., 1992, Pasteurella haemolytica leukotoxin inhibits mitogen-induced bovine peripheral blood mononuclear cell proliferation in vitro. Microb. Pathog. 12: 459–463.Google Scholar
  27. Devenish, J., Brown. J.E., and Rosendal, S., 1992. Association of the RTX proteins of Actinobacillus pleuropneumoniae with hemolytic, CAMP. and neutrophil-cytotoxic activities, Infect. Immun. 60: 2139–2142.Google Scholar
  28. Devenish, J., Rosendal, S. Johnson, R., and Hubler, S., 1989, Tmmunoserological comparison of 104kilodalton proteins associated with hemolysis and cytolysis in Actinobacillus pleuropneumoniae, actinobacillus suis, Pasteurella haemolytica, and Escherichia coli, Infect. Immun. 57: 3210–3213.Google Scholar
  29. Dom. P., Haesebrouck, F., Kamp, E.M., and Smits, M.A., 1992. Influence of Actinobacillus pleuropneumoniae serotype 2 and its cytolysins on porcine neutrophil chemiluminescence. Infect. Immun. 60: 4328–4334.PubMedGoogle Scholar
  30. Dyer. R.M., Benson, C.E., and Boy, M.G., 1984. Production of superoxide anion by bovine pulmonary macrophages challenged with soluble and particulate stimuli, Am. J. Vet. Res. 46: 336–341.Google Scholar
  31. Ermert, L., Rousseau, S., Schulte, H. Birkemeyer, R.G., Grimminger, Ff., Bhakdi, S., Duncker. H.R., and Seeger, W., 1992, Induction of severe vascular leakage by low doses of Escherichia coli hemolysin in perfused rabbit lungs. Lab. Invest. 66: 362.Google Scholar
  32. Gadeberg, O.V., and Orskov, I. 1984. in vitro cytotoxic effect of à-hemolytic Escherichia coli on human blood granulocytes. Infect. Immun. 45: 255–260.Google Scholar
  33. Grimminger, F., Walmrath, D., Birkemeyer, R.G., Bhakdi, S., and Seeger, W., 1990, Leukotriene and hydroxyeicosatetraenoic acid generation elicited by low doses of Escherichia coli hemolysin in rabbit lungs, Infect. Immun. 58: 2659–2663.PubMedGoogle Scholar
  34. Henricks, P.A., Binkhorst, G.J., Drijver. A.A., and Nijkamp, F.P., 1992, Pasteurella haemolytica leukotoxin enhances production of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by bovine polymorphonuclear leukocytes, Infect. Immun. 60: 3238–3243.Google Scholar
  35. Inzana, T.J., 1991, Virulence properties of Actinohacillus pleuropneumoniae, Microb. Pathog. 11:305–316. Iwase, M., Korchak, H.M„ Lally. E.T., Berthold. P., and Taichman, N.S., 1992, Lytic effects of Actinobacillus actinomycetemcomitans leukotoxin on human neutrophil cytoplasts,/. Leukoc. Biol. 52: 224–227.Google Scholar
  36. Iwase, M. Lally, E.T., Berthold. P. Korchak. H.M., and Taichman, N.S., 1990, Effects of cations and osmotic pmtectants on cytolytic activity of Actinohacillus actinomycetemcomitans leukotoxin, Infect. Immun. 58: 1782–1788.PubMedGoogle Scholar
  37. Jonas. D. Schultheis, B. Klas. C., Krammer. P.H. and Bhakdi. S., 1993. Cytocidal effects of Escherichia coli hemolysin on human T lymphocytes, Infect. Immun. 61: 1715–1721.PubMedGoogle Scholar
  38. Kaehler. K.L., Markham. R.J.F., Muscoplat, C.C., and Johnson, D.W., 1980a, Evidence of cytocidal effects of Pasteurella haemolytica on bovine peripheral blood mononuclear leukocytes, Am.1. Vet. Res. 41: 1690–1693.Google Scholar
  39. Koehler, K,L., Markham. R.J.F., Muscoplat. C.C., and Johnson, D.W., 1980b, Evidence of species specificity in the cytocidal effects of Pasteurella haemolytica, Infect. Immun. 30: 615–616.PubMedGoogle Scholar
  40. Kolodrubetz, D., Dailey, T., Ebersole. J., and Kraig, E., 1989, Cloning and expression of the leukotoxin gene from Actinobacillus actinomycetemcomitans, Infect. Immun. 57:1465–1469.Google Scholar
  41. Kraig. E., Dailey, T. and Kolodrubetz, D. 1990. Nucleotide sequence of the leukotoxin gene from Actinobacillus actinomycetemcomitans: homology to the alpha-hemolysin/leukotoxin gene family, Infect. immun. 58: 920–929.PubMedGoogle Scholar
  42. Lalonde. G., McDonald. T.V., Gardner, P. and O’Hanley, P.D., 1989, Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers, J. Biol. Chem. 264: I3559–13564.Google Scholar
  43. Maheswaran, S.K., Berggren, K.A., Simonson. R.R., Ward, G.E., and Muscoplat, C.C., 1980, Kinetics of interaction and fate of Pasteurella hemolytica: bovine alveolar macrophages, Infect. Immun. 30: 254262.Google Scholar
  44. Maheswaran, S.K., Kansan, M.S., Weiss. D.J., Reddy. K.R., Townsend, E.L., Yoo, H.S., Lee, B.W., and Whiteley, L.O., 1993, Enhancement of neutrophil-mediated injury to bovine pulmonary endothelial cells by Pasteurella haemolytica leukotoxin, Infect. Immun. 61: 2618–2625.Google Scholar
  45. Majury, A.L., and Shewen, P.E., 1991, The effect of Pasteurella haemolytica Al leukotoxic culture supernate on the in vitro proliferative response of bovine lymphocytes, Vet. Immunol. Immunopathol. 29: 41–56.Google Scholar
  46. Mangan, D.F., Taichman, N.S., Lally, E.T., and Wahl, S.M., 1991, Lethal effects of Actinobacillus actinomycetemcomitans leukotoxin on human T lymphocytes, Infect. Immun. 59: 3267–3272.PubMedGoogle Scholar
  47. Markham, R.J.F., Ramnarine, M.C.R., and Muscoplat. C.C., 1982, Cytotoxic effect of Pasteurella haemolytica on bovine polymorphonuclear leukocytes and impaired production of chemotactic factors by Pasteurella haemolytica-infected alveolar macrophages, Am. J. Vet. Res. 43: 285–288.Google Scholar
  48. Menestrina, G., Moser, C., Pellet, S., and Welch. R., 1994, Pore-formation by Escherichia coli hemolysin ( Hly A) and other members of the RTX toxins family, Toxicology 87: 249–267.Google Scholar
  49. Mosier, D.A., Confer, A.W., and Panciera, R.J., 1989, The evaluation of vaccines for bovine pneumonic pasteurellosis, Res. Vet. Sci. 47: 1–10.PubMedGoogle Scholar
  50. Mutters, R., Mannheim. W., and Bisgaard, M., 1989, Taxonomy of the group, in: Pasteurella and Pasteurellosis, C. Adlam and J.M. Rutter. ed., Academic Press, NY.Google Scholar
  51. Nicolet, J., 1990, Overview of the virulence attributes of the HAP-group of bacteria, Can. J. Vet. Res. 54:S12-S 15.Google Scholar
  52. Ohta, H., Hara, H., Fukui. K. Kurihara, H., Murayama, Y. and Kato, K., 1993, Association of Actinobacillus actinomycetemcomitans leukotoxin with nucleic acids on the bacterial cell surface. Infect. Immn. 61: 4878–4884.Google Scholar
  53. Ohta, H. Kato, K., Kokeguchi, S., Hara, H., Fukui, K., and Murayama, Y., 1991, Nuclease-sensitive binding of an Actinobacillus actinomycetemcomitans leukotoxin to the bacterial cell surface, Infect. Immun. 59: 4599–4605.Google Scholar
  54. Ortiz-Carranza, O., and Czuprynski. C.J., 1992, Activation of bovine neutrophils by Pasteurella haemolytica leukotoxin is calcium dependent, J. Leukoc. Biol. 52: 558–564.Google Scholar
  55. Ostolaza, H., Bartolome, B. Serra, J.L., de la Cruz. F., and Goni, F.M., 1991, a-Hemolysin from E. coli. Purification and self-aggregation properties, FEBS Lett. 280: 195–198.Google Scholar
  56. Pace, L.W., Kreeger, J.M., Bailey, K.L., Turnquist, S.E., and Fales, W.H., 1993, Serum levels of tumor necrosis factor-a in calves experimentally infected with Pasteurella haemolytica Al. Vet. Immnol. Immunopathol. 35: 353–364.Google Scholar
  57. Rabie, G., Lally, E.T., and Shenker, B.J., 1988, immunosuppressive properties of Actinobacillus actinomycetemcomitans leukotoxin, Infect. Immun. 56: 122–127.Google Scholar
  58. Sebo, P., and Ladant, D., 1993, Repeat sequences in the Bordetella pertussis adenylate cyclase toxin can be recognized as alternative carboxy-proximal secretion signals by the Escherichia coli a-haemolysin translocator, Mol. Microbiol. 9: 999–1009.PubMedCrossRefGoogle Scholar
  59. Seeger. W. Oberritz, R., Thomas, M., Walmrath, D., Sutton), N., Holland LB., Grimminger, F. Eberspacher, B., Hugo, F., and Bhakdi, S., 1991, Lung vascular injury after administration of viable hemolysinforming Escherichia coli in isolated rabbit lungs, Am. Rev. Respir. Dis. 143: 797–805.Google Scholar
  60. Sharma, S.A., Olchowy, T.W.J., and Breider, M.A., 1991, Alveolar macrophage and neutrophil interactions in Pasteurella haemolytica-induced endothelial cell injury, J. Infect. Dis. 165: 651–657.CrossRefGoogle Scholar
  61. Shewen. P.E., and Wilkie. B.N., 1982, Cytotoxin of Pasteurella haemolytica acting on bovine leukocytes, Infect. Immun. 35: 91–94.PubMedGoogle Scholar
  62. Skoyen, S., Czuprynski, C.J., Saban, R. 1993, Effects of the Pasteurella haemolytica leukotoxin and LPS on bovine lung parenchymal tissue in vitro, 74th Annu. Conf. Res. Work. Anim. Dis., Abst. P123.Google Scholar
  63. Slocombe, R.F., Malark, F., Ingersoll. R., Derksen, F.J., and Robinson, N.E., 1985, The importance of neutrophils in the pathogenesis of acute pneumonic pasteurellosis, Am. J. Vet. Res. 46: 2253–2258.Google Scholar
  64. Stanley. P.L.D., Diaz, P., Bailey, M.J.A., Gygi, D., Juarez. A., and Hughes, C., 1993, Loss of activity in the secreted form of Escherichia coli haemolysin caused by an rfaP lesion in core lipopolysaccharide assembly, Mol. Microbiol. 10: 781–787.PubMedCrossRefGoogle Scholar
  65. Stevens, P., and Czuprynski, C., 1995, Dissociation of cytolysis and monokine release by bovine mononuclear phagocytes incubated with Pasteurella haemolytica partially-purified leukotoxin and lipopolysaccharide. Can. J. Vet. Res. (in press, April).Google Scholar
  66. Strathdee, C.A., and Lo, R.Y.C., 1987, Extensive homology between the leukotoxin of Pasteurella haemolytica Al and the alpha-hemolysin of Escherichia coli. Infect. Immun. 55: 3233–3236.Google Scholar
  67. Styrt, B., Walker, R.D., Dahl, L.D., and Potter A., 1990, Time and temperature dependence of granulocyte damage by leukotoxic supernatants from Pasteurella haemolytica Al, J. Gen. Microbiol. 136: 2173 2178.Google Scholar
  68. Sutherland, A.D., Effects of Pasteurella haemolytica cytotoxin on ovine peripheral blood leucocytes and lymphocytes obtained from gastric lymph. Vet. Microbiol. 10: 431–438.Google Scholar
  69. Suttorp, N., Fuhrmann, M. Tannert-Otto. S. Grimminger, F., and Bhakdi, S., 1993, Pore-forming bacterial toxins potently induce release of nitric oxide in porcine endothelial cells, J. Exp. Med. 178: 337–341.Google Scholar
  70. Taichman, N.S., Iwase, M., Lally, E.T., Shattil, S.J., Cunningham, M.E., and Korchak, H.M., 1991. Early changes in cytosolic calcium and membrane potential induced by Actinohacillus actinomycetemcomitans leukotoxin in susceptible and resistant target cells, J. Immunol. 147: 35873594.Google Scholar
  71. Tsai, C.-C. McArthur. W.P., Baehni, P.C., Hammond, B.F., and Taichman, N.S., 1979. Extraction and partial characterization of a leukotoxin from a plaque-derived gram-negative microorganism, Infect. Immun. 25: 427–439.PubMedGoogle Scholar
  72. Tsai, C.-C., Shenker. B.J., DiRienzo, J.M., Malamud. D., and Taichman, N.S., 1984, Extraction and isolation of leukotoxin from Actinobacillus actinomycetemcomitans with polymyxin B, Infect. Immun. 43: 700705.Google Scholar
  73. Udeze, F.A., and Kadis, S., 1992, Effects of Actinohacillns pleuropneumoniae hemolysin on porcine neutrophil function, Infect. Immun. 60: 1558–1567.PubMedGoogle Scholar
  74. van Leengoed, L.A.M.G., and Dickerson, H.W., 1992, influence of calcium on secretion and activity of the cytolysins of Actinobacilius pleuropneumoniae. Infect. Immun. 60:353–359.Google Scholar
  75. Walker. R.D., Corstvet, R.E. and Panciera, R.J., 1979, Study of bovine pulmonary response to Pasteurella haemolytica: Pulmonary macrophage response, Am. J. Vet. Res. 41: 1008–1014.Google Scholar
  76. Wandersman. C., and Letoffe. S., 1993, involvent of lipopolysaccharide in the secretion of Escherichia coli a-haemolysin and Erwinia chrysanthemi proteases, Molec. Microbiol. 7: 141–150.Google Scholar
  77. Weiss, D.I. Bauer, M.C., Whiteley, L.O., Maheswaran, S.K., Ames, T.R., 1991, Changes in blood and bronchoalveolar lavage fluid components in calves with experimentally induced pneumonic pasteurellosis, Am.I. Vet. Res. 52: 337–343.Google Scholar
  78. Welch, R.A., Forestier, C., Lobo. A., Pellett, S., Thomas Jr. W., and Rowe, G. 1992, The synthesis and function of the Escherichia coli hemolysin and related RTX exotoxins, Microbiol. Immunol. 105: 2936.Google Scholar

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© Springer Science+Business Media New York 1995

Authors and Affiliations

  • C. J. Czuprynski
    • 1
  1. 1.Dept. Pathobiological Sciences School of Veterinary MedicineUniversity of WisconsinMadisonUSA

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