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Infect Immun. 1996 July; 64(7): 2687–2694.
PMCID: PMC174127

Pasteurella haemolytica leukotoxin induces bovine leukocytes to undergo morphologic changes consistent with apoptosis in vitro.

Abstract

Infection of the bovine lung with Pasteurella haemolytica results in an acute respiratory disorder known as pneumonic pasteurellosis. One of the key virulence determinants used by this bacterium is secretion of an exotoxin that is specific for ruminant leukocytes (leukotoxin). At low concentrations, the leukotoxin can activate ruminant leukocytes, whereas at higher concentrations, it inhibits leukocyte functions and is cytolytic, presumably as a result of pore formation and subsequent membrane permeabilization. We have investigated the possibility that the activation-inhibition paradox is explained in part by leukotoxin-mediated apoptosis (i.e., activation-induced cell death) of bovine leukocytes. Incubation of bovine leukocytes with P. haemolytica leukotoxin caused marked cytoplasmic membrane blebbing (zeiosis) and chromatin condensation and margination, both of which are hallmarks of apoptosis. The observed morphologic changes in bovine leukocytes were leukotoxin dependent, because they were significantly diminished in the presence of an anti-leukotoxin monoclonal antibody. In addition, bovine leukocytes incubated with culture supernatant from a mutant strain of P. haemolytica that does not produce any detectable leukotoxin failed to exhibit the morphologic changes characteristic of cells undergoing apoptosis. These observations may represent an important mechanism by which P. haemolytica overwhelms host defenses, contributing to the fibrinous pleuropneumonia characteristic of bovine pasteurellosis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Baluyut CS, Simonson RR, Bemrick WJ, Maheswaran SK. Interaction of Pasteurella haemolytica with bovine neutrophils: identification and partial characterization of a cytotoxin. Am J Vet Res. 1981 Nov;42(11):1920–1926. [PubMed]
  • Beaver JP, Waring P. Lack of correlation between early intracellular calcium ion rises and the onset of apoptosis in thymocytes. Immunol Cell Biol. 1994 Dec;72(6):489–499. [PubMed]
  • Bertrand R, Solary E, O'Connor P, Kohn KW, Pommier Y. Induction of a common pathway of apoptosis by staurosporine. Exp Cell Res. 1994 Apr;211(2):314–321. [PubMed]
  • Bhakdi S, Mackman N, Nicaud JM, Holland IB. Escherichia coli hemolysin may damage target cell membranes by generating transmembrane pores. Infect Immun. 1986 Apr;52(1):63–69. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Boehm DF, Welch RA, Snyder IS. Domains of Escherichia coli hemolysin (HlyA) involved in binding of calcium and erythrocyte membranes. Infect Immun. 1990 Jun;58(6):1959–1964. [PMC free article] [PubMed]
  • Bozdech MJ, Bainton DF. Identification of alpha-naphthyl butyrate esterase as a plasma membrane ectoenzyme of monocytes and as a discrete intracellular membrane-bounded organelle in lymphocytes. J Exp Med. 1981 Jan 1;153(1):182–195. [PMC free article] [PubMed]
  • Chang YF, Young R, Post D, Struck DK. Identification and characterization of the Pasteurella haemolytica leukotoxin. Infect Immun. 1987 Oct;55(10):2348–2354. [PMC free article] [PubMed]
  • Chidambaram M, Sharma B, Petras SF, Reese CP, Froshauer S, Weinstock GM. Isolation of Pasteurella haemolytica leukotoxin mutants. Infect Immun. 1995 Mar;63(3):1027–1032. [PMC free article] [PubMed]
  • Clinkenbeard KD, Clarke CR, Hague CM, Clinkenbeard P, Srikumaran S, Morton RJ. Pasteurella haemolytica leukotoxin-induced synthesis of eicosanoids by bovine neutrophils in vitro. J Leukoc Biol. 1994 Nov;56(5):644–649. [PubMed]
  • Clinkenbeard KD, Mosier DA, Confer AW. Transmembrane pore size and role of cell swelling in cytotoxicity caused by Pasteurella haemolytica leukotoxin. Infect Immun. 1989 Feb;57(2):420–425. [PMC free article] [PubMed]
  • Clinkenbeard KD, Upton ML. Lysis of bovine platelets by Pasteurella haemolytica leukotoxin. Am J Vet Res. 1991 Mar;52(3):453–457. [PubMed]
  • Cohen JJ, Duke RC, Fadok VA, Sellins KS. Apoptosis and programmed cell death in immunity. Annu Rev Immunol. 1992;10:267–293. [PubMed]
  • Confer AW, Panciera RJ, Clinkenbeard KD, Mosier DA. Molecular aspects of virulence of Pasteurella haemolytica. Can J Vet Res. 1990 Apr;54 (Suppl):S48–S52. [PubMed]
  • Czuprynski CJ, Hamilton HL, Noel EJ. Ingestion and killing of Pasteurella haemolytica A1 by bovine neutrophils in vitro. Vet Microbiol. 1987 May;14(1):61–74. [PubMed]
  • Czuprynski CJ, Noel EJ, Ortiz-Carranza O, Srikumaran S. Activation of bovine neutrophils by partially purified Pasteurella haemolytica leukotoxin. Infect Immun. 1991 Sep;59(9):3126–3133. [PMC free article] [PubMed]
  • Czuprynski CJ, Ortiz-Carranza O. Pasteurella haemolytica leukotoxin inhibits mitogen-induced bovine peripheral blood mononuclear cell proliferation in vitro. Microb Pathog. 1992 Jun;12(6):459–463. [PubMed]
  • Dom P, Haesebrouck F, Kamp EM, Smits MA. NAD-independent Actinobacillus pleuropneumoniae strains: production of RTX toxins and interactions with porcine phagocytes. Vet Microbiol. 1994 Apr;39(3-4):205–218. [PubMed]
  • Dowd DR, Miesfeld RL. Evidence that glucocorticoid- and cyclic AMP-induced apoptotic pathways in lymphocytes share distal events. Mol Cell Biol. 1992 Aug;12(8):3600–3608. [PMC free article] [PubMed]
  • Ermert L, Rousseau S, Schütte H, Birkemeyer RG, Grimminger F, Bhakdi S, Duncker HR, Seeger W. Induction of severe vascular leakage by low doses of Escherichia coli hemolysin in perfused rabbit lungs. Lab Invest. 1992 Mar;66(3):362–369. [PubMed]
  • Furuya Y, Lundmo P, Short AD, Gill DL, Isaacs JT. The role of calcium, pH, and cell proliferation in the programmed (apoptotic) death of androgen-independent prostatic cancer cells induced by thapsigargin. Cancer Res. 1994 Dec 1;54(23):6167–6175. [PubMed]
  • Gentry MJ, Srikumaran S. Neutralizing monoclonal antibodies to Pasteurella haemolytica leukotoxin affinity-purify the toxin from crude culture supernatants. Microb Pathog. 1991 May;10(5):411–417. [PubMed]
  • Gerbig DG, Jr, Walker RD, Baker JC, Foster JS, Moore RN. Calcium ion involvement in the action of Pasteurella haemolytica leukotoxin. Vet Microbiol. 1989 Apr;19(4):325–335. [PubMed]
  • Grimminger F, Thomas M, Obernitz R, Walmrath D, Bhakdi S, Seeger W. Inflammatory lipid mediator generation elicited by viable hemolysin-forming Escherichia coli in lung vasculature. J Exp Med. 1990 Oct 1;172(4):1115–1125. [PMC free article] [PubMed]
  • Henricks PA, Binkhorst GJ, Drijver AA, Nijkamp FP. Pasteurella haemolytica leukotoxin enhances production of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by bovine polymorphonuclear leukocytes. Infect Immun. 1992 Aug;60(8):3238–3243. [PMC free article] [PubMed]
  • Himmel ME, Yates MD, Lauerman LH, Squire PG. Purification and partial characterization of a macrophage cytotoxin from Pasteurella haemolytica. Am J Vet Res. 1982 May;43(5):764–767. [PubMed]
  • Hogquist KA, Nett MA, Unanue ER, Chaplin DD. Interleukin 1 is processed and released during apoptosis. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8485–8489. [PubMed]
  • Hughes HP, Campos M, McDougall L, Beskorwayne TK, Potter AA, Babiuk LA. Regulation of major histocompatibility complex class II expression by Pasteurella haemolytica leukotoxin. Infect Immun. 1994 May;62(5):1609–1615. [PMC free article] [PubMed]
  • Jonas D, Schultheis B, Klas C, Krammer PH, Bhakdi S. Cytocidal effects of Escherichia coli hemolysin on human T lymphocytes. Infect Immun. 1993 May;61(5):1715–1721. [PMC free article] [PubMed]
  • Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972 Aug;26(4):239–257. [PMC free article] [PubMed]
  • Khelef N, Zychlinsky A, Guiso N. Bordetella pertussis induces apoptosis in macrophages: role of adenylate cyclase-hemolysin. Infect Immun. 1993 Oct;61(10):4064–4071. [PMC free article] [PubMed]
  • Kluck RM, McDougall CA, Harmon BV, Halliday JW. Calcium chelators induce apoptosis--evidence that raised intracellular ionised calcium is not essential for apoptosis. Biochim Biophys Acta. 1994 Sep 8;1223(2):247–254. [PubMed]
  • König B, König W, Scheffer J, Hacker J, Goebel W. Role of Escherichia coli alpha-hemolysin and bacterial adherence in infection: requirement for release of inflammatory mediators from granulocytes and mast cells. Infect Immun. 1986 Dec;54(3):886–892. [PMC free article] [PubMed]
  • König B, Ludwig A, Goebel W, König W. Pore formation by the Escherichia coli alpha-hemolysin: role for mediator release from human inflammatory cells. Infect Immun. 1994 Oct;62(10):4611–4617. [PMC free article] [PubMed]
  • Lee KS, Tsien RW. Mechanism of calcium channel blockade by verapamil, D600, diltiazem and nitrendipine in single dialysed heart cells. Nature. 1983 Apr 28;302(5911):790–794. [PubMed]
  • Lefkowitz RJ. Direct binding studies of adrenergic receptors: biochemical, physiologic, and clinical implications. Ann Intern Med. 1979 Sep;91(3):450–458. [PubMed]
  • Ludwig A, Jarchau T, Benz R, Goebel W. The repeat domain of Escherichia coli haemolysin (HlyA) is responsible for its Ca2+-dependent binding to erythrocytes. Mol Gen Genet. 1988 Nov;214(3):553–561. [PubMed]
  • Maheswaran SK, Kannan MS, Weiss DJ, Reddy KR, Townsend EL, Yoo HS, Lee BW, Whiteley LO. Enhancement of neutrophil-mediated injury to bovine pulmonary endothelial cells by Pasteurella haemolytica leukotoxin. Infect Immun. 1993 Jun;61(6):2618–2625. [PMC free article] [PubMed]
  • Maheswaran SK, Weiss DJ, Kannan MS, Townsend EL, Reddy KR, Whiteley LO, Srikumaran S. Effects of Pasteurella haemolytica A1 leukotoxin on bovine neutrophils: degranulation and generation of oxygen-derived free radicals. Vet Immunol Immunopathol. 1992 Jun;33(1-2):51–68. [PubMed]
  • Majury AL, Shewen PE. The effect of Pasteurella haemolytica A1 leukotoxic culture supernate on the in vitro proliferative response of bovine lymphocytes. Vet Immunol Immunopathol. 1991 Aug;29(1-2):41–56. [PubMed]
  • Mangan DF, Taichman NS, Lally ET, Wahl SM. Lethal effects of Actinobacillus actinomycetemcomitans leukotoxin on human T lymphocytes. Infect Immun. 1991 Sep;59(9):3267–3272. [PMC free article] [PubMed]
  • Menestrina G, Mackman N, Holland IB, Bhakdi S. Escherichia coli haemolysin forms voltage-dependent ion channels in lipid membranes. Biochim Biophys Acta. 1987 Nov 27;905(1):109–117. [PubMed]
  • Ondrias K, Stasko A, Jancinová V, Balgavý P. Comparison of the effect of eleven beta-adrenoceptor blocking drugs in perturbing lipid membrane: an ESR spectroscopy study. Mol Pharmacol. 1987 Jan;31(1):97–102. [PubMed]
  • Ortiz-Carranza O, Czuprynski CJ. Activation of bovine neutrophils by Pasteurella haemolytica leukotoxin is calcium dependent. J Leukoc Biol. 1992 Nov;52(5):558–564. [PubMed]
  • Oshimi Y, Miyazaki S. Fas antigen-mediated DNA fragmentation and apoptotic morphologic changes are regulated by elevated cytosolic Ca2+ level. J Immunol. 1995 Jan 15;154(2):599–609. [PubMed]
  • Petras SF, Chidambaram M, Illyes EF, Froshauer S, Weinstock GM, Reese CP. Antigenic and virulence properties of Pasteurella haemolytica leukotoxin mutants. Infect Immun. 1995 Mar;63(3):1033–1039. [PMC free article] [PubMed]
  • Rogel A, Hanski E. Distinct steps in the penetration of adenylate cyclase toxin of Bordetella pertussis into sheep erythrocytes. Translocation of the toxin across the membrane. J Biol Chem. 1992 Nov 5;267(31):22599–22605. [PubMed]
  • Sarih M, Souvannavong V, Brown SC, Adam A. Silica induces apoptosis in macrophages and the release of interleukin-1 alpha and interleukin-1 beta. J Leukoc Biol. 1993 Nov;54(5):407–413. [PubMed]
  • Scales B, McIntosh DA. Effects of propranolol and its optical isomers on the radiocalcium uptake and the adenosine triphosphatases of skeletal and cardiac sarcoplasmic reticulum fractions (SRF). J Pharmacol Exp Ther. 1968 Apr;160(2):261–267. [PubMed]
  • Searle J, Kerr JF, Bishop CJ. Necrosis and apoptosis: distinct modes of cell death with fundamentally different significance. Pathol Annu. 1982;17(Pt 2):229–259. [PubMed]
  • Sharma SA, Olchowy TW, Breider MA. Alveolar macrophage and neutrophil interactions in Pasteurella haemolytica-induced endothelial cell injury. J Infect Dis. 1992 Apr;165(4):651–657. [PubMed]
  • Shenker BJ, Vitale LA, Keiba I, Harrison G, Berthold P, Golub E, Lally ET. Flow cytometric analysis of the cytotoxic effects of Actinobacillus actinomycetemcomitans leukotoxin on human natural killer cells. J Leukoc Biol. 1994 Feb;55(2):153–160. [PubMed]
  • Shewen PE, Wilkie BN. Cytotoxin of Pasteurella haemolytica acting on bovine leukocytes. Infect Immun. 1982 Jan;35(1):91–94. [PMC free article] [PubMed]
  • Shewen PE, Wilkie BN. Evidence for the Pasteurella haemolytica cytotoxin as a product of actively growing bacteria. Am J Vet Res. 1985 May;46(5):1212–1214. [PubMed]
  • Slocombe RF, Malark J, Ingersoll R, Derksen FJ, Robinson NE. Importance of neutrophils in the pathogenesis of acute pneumonic pasteurellosis in calves. Am J Vet Res. 1985 Nov;46(11):2253–2258. [PubMed]
  • Stevens P, Czuprynski C. Dissociation of cytolysis and monokine release by bovine mononuclear phagocytes incubated with Pasteurella haemolytica partially purified leukotoxin and lipopolysaccharide. Can J Vet Res. 1995 Apr;59(2):110–117. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Takei N, Endo Y. Ca2+ ionophore-induced apoptosis on cultured embryonic rat cortical neurons. Brain Res. 1994 Jul 25;652(1):65–70. [PubMed]
  • Yoo HS, Rajagopal BS, Maheswaran SK, Ames TR. Purified Pasteurella haemolytica leukotoxin induces expression of inflammatory cytokines from bovine alveolar macrophages. Microb Pathog. 1995 Apr;18(4):237–252. [PubMed]
  • Weksler BB, Gillick M, Pink J. Effect of propranolol on platelet function. Blood. 1977 Feb;49(2):185–196. [PubMed]
  • Welch RA. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol. 1991 Mar;5(3):521–528. [PubMed]
  • Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. [PubMed]
  • Zychlinsky A, Fitting C, Cavaillon JM, Sansonetti PJ. Interleukin 1 is released by murine macrophages during apoptosis induced by Shigella flexneri. J Clin Invest. 1994 Sep;94(3):1328–1332. [PMC free article] [PubMed]

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