1887

Abstract

Mutants of strain HK 361 (serotype 2) were isolated which were deficient in type II (Ca-dependent) haemolysin activity (Hly). Some of the Hly mutants secreted a potent, heat-labile extracellular cytotoxic activity against porcine alveolar macrophages. Comparison of cell-free culture supernatant from the parent strain and some Hly mutants by SDS-PAGE and immunoblotting revealed the loss of a major extracellular polypeptide of 109 kDa. Two Hly mutants which in addition failed to secrete a 120 kDa polypeptide produced no extracellular cytotoxic activity, suggesting that the 120 kDa protein was the cytotoxin. Antiserum raised to the culture supernatant from a Hly mutant lacking the 109 kDa polypeptide recognized the 120 kDa band, but not the 109 kDa band, in immunoblots and neutralized the cytotoxic activity, but not the haemolytic activity, of . The 120 kDa polypeptide and extracellular cytotoxic activity were widespread among strains, but absent from related bacterial pathogens of the pig: and . A clear correlation was found between the presence of the 120 kDa polypeptide and cytotoxic activity in culture supernatants. The cytotoxic activity of all the strains tested was neutralized by antibody to the Hly extracellular material and by convalescent pig serum. It is proposed that the 120 kDa polypeptide represents the cytotoxin of , that it is distinct from the haemolysin, and that it be termed pleurotoxin.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-3-561
1991-03-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/3/mic-137-3-561.html?itemId=/content/journal/micro/10.1099/00221287-137-3-561&mimeType=html&fmt=ahah

References

  1. Bendixen P. H., Shewen P. E., Rosendal S., Wilkie B. N. 1981; Toxicity of Haemophilus pleuropneumoniae for porcine lung macrophages, peripheral blood monocytes, and testicular cells. Infection and Immunity 33:673–676
    [Google Scholar]
  2. Bertram T. A. 1988; Pathobiology of acute pulmonary lesions in swine infected with Haemophilus Actinobacillus pleuropneumoniae. Canadian Veterinary Journal 29:574–577
    [Google Scholar]
  3. Chang Y.-F., Young R., Post D., Struck D. K. 1987; Identification and characterisation of the Pasteurella haemolytica leukotoxin. Infection and Immunity 55:2348–2354
    [Google Scholar]
  4. Chang Y.-F., Young R., Struck D. K. 1989; Cloning and characterization of a hemolysin gene from Actinobacillus Haemophilus pleuropneumoniae. DNA 8:635–647
    [Google Scholar]
  5. Davis B., Barsoum I. 1980; Human alveolar macrophages: effect of endotoxin in vitro. Infection and Immunity 30:753–758
    [Google Scholar]
  6. Devenish J., Rosendal S. 1989; Identification of the heat-labile hemolysin of Actinobacillus pleuropneumoniae serotype 1. Canadian Journal of Veterinary Research 53:251–254
    [Google Scholar]
  7. Devenish J., Rosendal S., Johnson R., Hubler S. 1989; Immunoserological comparison of 104-kilodalton proteins associated with hemolysis and cytolysis in Actinobacillus pleuropneumoniae, Actinobacillus suis, Pasteur ella haemolytica, and Escherichia coli. Infection and Immunity 57:3210–3213
    [Google Scholar]
  8. Fedorka-Cray P. J., Huether M. J., Stine D. L., Anderson G. A. 1990; Efficacy of a cell extract from Actinobacillus (Haemophilus) pleuropneumoniae serotype 1 against disease in swine. Infection and Immunity 58:358–365
    [Google Scholar]
  9. Frey J., Nicolet J. 1988e; Purification and partial characterisation of a hemolysin produced by Actinobacillus pleuropneumoniae type strain 4074. FEMS Microbiology Letters 55:41–46
    [Google Scholar]
  10. Frey J., Nicolet J. 1988b; Regulation of hemolysin expression in Actinobacillus pleuropneumoniae serotype 1 by Ca2+. Infection and Immunity 56:2570–2575
    [Google Scholar]
  11. Frey J., Perrin J., Nicolet J. 1989; Cloning and expression of a cohemolysin, the CAMP factor of Actinobacillus pleuropneumoniae. Infection and Immunity 57:2050–2056
    [Google Scholar]
  12. Gulig P. A., Frisch C. F., Hansen E. J. 1983; A set of two monoclonal antibodies specific for the cell surface-exposed 39K major outer membrane protein of Haemophilus influenzae type b defines all strains of this pathogen. Infection and Immunity 42:516–524
    [Google Scholar]
  13. Gulig P. A., McCracken G. H., Holmans P. L., Hansen E. J. 1984; Immunogenic proteins in cell-free culture supernatants of Haemophilus influenzae type b. Infection and Immunity 44:41–48
    [Google Scholar]
  14. Gygi D., Nicolet J., Frey J., Cross M., Koronakis V., Hughes C. 1990; Isolation of the Actinobacillus pleuropneumoniae haemolysin gene and the activation and secretion of the prohaemolysin by the HlyC, HlyB and HlyD proteins of Escherichia coli. Molecular Microbiology 4:123–128
    [Google Scholar]
  15. Harlow E., Lane D. 1988 Antibodies: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  16. Kamp E. M., Van Leengoed L. A. M. G. 1989; Serotype-related differences in production and type of heat-labile hemolysin and heat-labile cytotoxin of Actinobacillus (Haemophilus) pleuropneumoniae. Journal of Clinical Microbiology 27:1187–1191
    [Google Scholar]
  17. Kilian M., Nicolet J., Biberstein E. L. 1978; Biochemical and serological characterization of Haemophilus pleuropneumoniae (Matthews and Pattison 1961) Shope 1964 and proposal of a neotype strain. International Journal of Systematic Bacteriology 28:20–26
    [Google Scholar]
  18. Kume K., Nakai T., Sawata A. 1986; Interaction between heat-stable hemolytic substance from Haemophilus pleuropneumoniae and porcine pulmonary macrophages in vitro. Infection and Immunity 51:563–570
    [Google Scholar]
  19. Leijh P. C. J., Van Furth R., Van Zwet T. L. 1986; In vitro determination of phagocytosis and intracellular killing by polymor-phonuclear and mononuclear phagocytes. Handbook of Experimental Immunology 2 Cellular Immunology, 4.46.1–46.21 Weir D. M. Oxford: Blackwell Scientific Publications;
    [Google Scholar]
  20. Liggett A. D., Harrison L. R., Farrell R. L. 1987; Sequential study of lesion development in experimental haemophilus pleuropneumonia. Research in Veterinary Science 42:204–221
    [Google Scholar]
  21. Ludwig A., Jarchau T., Benz R., Goebel W. 1988; The repeat domain of E. coli hemolysin (HlyA) is responsible for its Ca2+-dependent binding to erythrocytes. Molecular and General Genetics 214:553–561
    [Google Scholar]
  22. Martin P. G., Lachance P., Niven D. F. 1985; Production of RNA-dependent haemolysin by Haemophilus pleuropneumoniae. Canadian Journal of Microbiology 31:456–462
    [Google Scholar]
  23. MILLER J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. Nicolet J. 1986; Haemophilus infections. In Diseases of Swine426–436 Leman A. D., Straw B., Glock R. D., Mengeling W. L., Penny R. H. C., Scholl E. Ames: Iowa State University Press;
    [Google Scholar]
  25. Nicolet J. 1990; Overview of the virulence attributes of the HAP-group of bacteria. Canadian Journal of Veterinary Research (Supplement) 54:S12–S15
    [Google Scholar]
  26. Rennie R. P., Freer J. H., Arbuthnott J. P. 1974; The kinetics of erythrocyte lysis by Escherichia coli haemolysin. Journal of Medical Microbiology 7:189–195
    [Google Scholar]
  27. Rosendal S., Mitchell W. R., Weber M., Wilson M. R., Zaman M. R. 1980; Haemophilus pleuropneumonia. Lung lesions induced by sonicated bacteria and sterile culture supernatant. Proceedings of the International Pig Veterinary Society Congress 6221
    [Google Scholar]
  28. Rosendal S., Devenish J., MacInnes J. I., Lumsden J. H., Watson S., Xun H. 1988; Evaluation of heat-sensitive, neutrophil-toxic, and hemolytic activity of Haemophilus (Actinobacillus) pleuropneumoniae. American Journal of Veterinary Research 49:1053–1058
    [Google Scholar]
  29. Rycroft A. N., Taylor D. J. 1987; Preparation and characterisation of envelope proteins from Haemophilus pleuropneumoniae. Veterinary Microbiology 15:303–314
    [Google Scholar]
  30. Sebunya T. N. K., Saunders J. R. 1983; Haemophilus pleuropneumoniae infection in swine: a review. Journal of the American Veterinary Medical Association 182:1331–1337
    [Google Scholar]
  31. Shope R. E. 1964; Porcine contagious pleuropneumonia. I. Exper-imental transmission, etiology and pathology. Journal of Experimental Medicine 119:357–368
    [Google Scholar]
  32. Strathdee C. A., Lo R. Y. C. 1987; Extensive homology between the leukotoxin of Pasteurella haemolytica and the a-hemolysin of Escherichia coli. Infection and Immunity 55:3233–3236
    [Google Scholar]
  33. Udeze F. A., Kadis S. 1988; Effect of Actinobacillus pleuropneumoniae hemolysin on viability and function of porcine phagocytes. Proceedings of the International Pig Veterinary Society Congress 1064
    [Google Scholar]
  34. Udezë F. A., Latimer K. S., Kadis S. 1987; Role of Haemophilus pleuropneumoniae lipopolysaccharide endotoxin in the pathogenesis of porcine Haemophilus pleuropneumonia. American Journal of Veterinary Research 48:768–773
    [Google Scholar]
  35. Van Leengoed L. A., Kamp E. M., Pol J. M. A. 1989; Toxicity of Haemophilus pleuropneumoniae to porcine lung macrophages. Veterinary Microbiology 19:337–349
    [Google Scholar]
  36. Weiss A. A., Falkow S. 1984; Genetic analysis of phase change in Bordetella pertussis. Infection and Immunity 43:263–269
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-3-561
Loading
/content/journal/micro/10.1099/00221287-137-3-561
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error