1887

Abstract

Bacterial exotoxins may contribute to the pathogenic potential of micro-organisms through interactions with cells of the host defence system as well as by directly damaging host tissue. The present studies were designed to explore mechanisms of interaction between bovine granulocytes and the leucotoxin produced by , a major cause of bovine respiratory disease. Leucotoxin-containing supernatant from A1 caused rapid cell death in isolated bovine granulocytes that was close to half-maximal by 5 min and nearly 90% complete after 30 min at 37 °C. Maintaining granulocytes at ice-water temperature markedly attenuated or prevented the toxic effect; furthermore, if exposed to supernatants at ice-water temperature and then washed, most cells remained viable even after rewarming to room temperature. However, even a very brief exposure (about 5 s) at 37 °C led to extensive cell death even after immediate cold dilution and washing. Granule enzymes such as arylsulphatase were released far more slowly than cytosol contents. Leucotoxin purified by column chromatography showed temperature dependence and divergence between cytosol and granule marker release similar to those observed with the crude supernatant preparation. These findings indicate that irreversible interaction between leucotoxin and bovine granulocytes is initiated very rapidly at 37 °C but markedly impeded at low temperature, while granule enzyme release follows cytosol marker release over a much longer period. The results suggest either a requirement for target cell metabolic activity to initiate toxin effects or a temperature-dependent receptor conformation, with granule enzyme release following as a secondary consequence of granulocyte death.

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1990-11-01
2021-10-27
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References

  1. Alouf J. E. 1980; Streptococcal toxins (streptolysin O, streptolysin S, erythrogenic toxin). Pharmacology and Therapeutics 11:661–717
    [Google Scholar]
  2. Alouf J. E., Loridan C. 1988; Production, purification and assay of streptolysin S. Methods in Enzymology 165:59–64
    [Google Scholar]
  3. Breider M. W., Walker R. D., Hopkins F. M., Schultz T. W., Bowersock T. L. 1988; Pulmonary lesions induced by Pasteurella haemolytica in neutrophil sufficient and neutrophil deficient calves. Canadian Journal of Veterinary Research 52:205–209
    [Google Scholar]
  4. Brown G. B., Roth J. A. 1990; Comparison of the response of bovine and human neutrophils to various stimuli. Veterinary Immunology and Immunopathology in the Press
    [Google Scholar]
  5. Chang Y. F., Renshaw H. W. 1986; Pasteurella haemolytica leukotoxin: comparison of 51chromium-release, trypan blue dye exclusion, and luminol-dependent chemiluminescence-inhibition assays for sensitivity in detecting leukotoxin activity. American Journal of Veterinary Research 47:134–138
    [Google Scholar]
  6. Chang Y. F., Renshaw H. W., Martens R. J., Livingston C. W. Jr 1986; Pasteurella haemolytica leukotoxin: chemiluminescent responses of peripheral blood leukocytes from several different mammalian species to leukotoxin- and opsonin-treated living and killed Pasteurella haemolytica and Staphylococcus aureus . American Journal of Veterinary Research 47:67–74
    [Google Scholar]
  7. Clinkenbeard K. D., Mosier D. A., Timko A. L., Confer A. W. 1989; Effects of Pasteurella haemolytica leukotoxin on cultured bovine lymphoma cells. American Journal of Veterinary Research 50:271–275
    [Google Scholar]
  8. Colton T. 1974 Statistics in Medicine Boston: Little, Brown & Co;
    [Google Scholar]
  9. Deneer H. G., Potter A. A. 1989; Iron-repressible outer-membrane proteins of Pasteurella haemolytica . Journal of General Microbiology 135:435–443
    [Google Scholar]
  10. Densen P., Mandell G. L. 1980; Phagocyte strategy vs. microbial tactics. Reviews of Infectious Diseases 2:817–836
    [Google Scholar]
  11. Emery D. L., Dufty J. H., Clark S. L. 1984; Biochemical and functional properties of a leucocidin produced by several strains of Fusobacterium necrophorum . Australian Veterinary Journal 61:382–387
    [Google Scholar]
  12. Emery D. L., Vaughan J. A. 1986; Generation of immunity against Fusobacterium necrophorum in mice inoculated with extracts containing leucocidin. Veterinary Microbiology 12:255–268
    [Google Scholar]
  13. Fluharty A. L., Edmond J. 1978; Arylsulfatases A and B from human liver. Methods in Enzymology 50:537–547
    [Google Scholar]
  14. Healy P. J. 1982; Lysosomal hydrolase activity in leucocytes from cattle, sheep, goats, horses and pigs. Research in Veterinary Science 33:275–279
    [Google Scholar]
  15. Kaehler K. L., Markam R. J. F., Muscoplat C. C., Johnson D. W. 1980; Evidence of cytocidal effects of Pasteurella haemolytica on bovine peripheral blood mononuclear leukocytes. American Journal of Veterinary Research 41:1690–1693
    [Google Scholar]
  16. Kanoe M., Yamamato T., Kai K., Blobel H. 1988; Effects of leukocidin from Fusobacterium necrophorum on bovine peripheral leukocytes in vitro . Zentralblatt für Bakteriologie und Hygiene A268:463–469
    [Google Scholar]
  17. Korzeniewski C., Callewaert D. M. 1983; An enzyme-release assay for natural cytotoxicity. Journal of Immunological Methods 64:313–320
    [Google Scholar]
  18. Lo R. Y. C., Strathdee C. A., Shewen P. A. 1987; Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1. Infection and Immunity 55:1987–1996
    [Google Scholar]
  19. Noda M., Hirayama T., Kato I., Matsuda F. 1980; Crystallization and properties of staphylococcal leukocidin. Biochimica et Biophysica Acta 633:33–44
    [Google Scholar]
  20. Noda M., Kato I. 1988; Purification and crystallization of staphylococcal leukocidin. Methods in Enzymology 165:22–33
    [Google Scholar]
  21. O’Brien J. K., Duffus W. P. H. 1987; Pasteurella haemolytica cytotoxin: relative susceptibility of bovine leukocytes. Veterinary Microbiology 13:321–334
    [Google Scholar]
  22. Potter A. A., Ready K., Gilchrist J. 1988; Purification of fimbriae from Pasteurella haemolytica A1. Microbial Pathogenesis 4:311–316
    [Google Scholar]
  23. Sklar L. A., Mcneil V. M., Jesaitis A. J., Painter R. G., Cochrane C. G. 1982; A continuous, spectroscopic analysis of the kinetics of elastase secretion by neutrophils. Journal of Biological Chemistry 257:5471–5475
    [Google Scholar]
  24. Slocombe R. F., Malark J., Ingersoll R., Derksen F. J., Robinson N. E. 1985; Importance of neutrophils in the pathogenesis of acute pneumonic pasteurellosis in calves. American Journal of Veterinary Research 46:2253–2258
    [Google Scholar]
  25. Strathdee C. A., Lo R. Y. C. 1987; Extensive homology between the leukotoxin of Pasteurella haemolytica A1 and the alpha-hemolysin of Escherichia coli . Infection and Immunity 55:3233–3236
    [Google Scholar]
  26. Styrt B., Johnson P. C., Klempner M. S. 1985; Differential lysis of plasma membranes and granules of human neutrophils by digitonin. Tissue & Cell 17:793–800
    [Google Scholar]
  27. Styrt B., Walker R. D., White J. C., Dahl L. D., Baker J. C. 1990; Granulocyte plasma membrane damage by leukotoxic supernatant from Pasteurella haemolytica A1 and protection by immune serum. Canadian Journal of Veterinary Research 54:146–150
    [Google Scholar]
  28. Walker R. D., Hopkins F. M., Schultz T. W., Mccracken M. D., Moore R. N. 1985; Changes in leukocyte populations in pulmonary lavage fluids of calves after inhalation of Pasteurella haemolytica . American Journal of Veterinary Research 46:2429–2433
    [Google Scholar]
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