1887

Abstract

Summary

Peritoneal polymorphonuclear neutrophils (PMN) from mice were tested for their ability to kill the yeast form of (Bd) and for their fungicidal mechanisms. PMN elicited from immune mice by the intraperitoneal injection of non-viable Bd (referred to as immunologically activated PMN or ActPMN) showed significantly enhanced fungicidal activity in comparison with PMN elicited with thioglycollate medium (ThioPMN) [means=44·7% (SD 12·8%) and 16·4% (SD 9·2%) killed; n= 14; p < 0·001]. Production of superoxide anion (O ) by ActPMN after stimulation with phorbol myristate acetate was enhanced in comparison with production by ThioPMN. Superoxide dismutase, which removes O , inhibited ActPMN killing by 75% (p < 0·001) when added to cultures immediately before challenge with Bd (optimal concentration : 6000 U/ml). Sodium azide, which inhibits myeloperoxidase and scavenges singlet oxygen (O), and catalase, which breaks down hydrogen peroxide (HO), inhibited ActPMN killing by 64% (p < 0·001) and 52% (p < 0·001), with optimal concentrations of 1 mM and 10 000 U/ml, respectively. Two agents that both scavenge O and antagonise hypochlorous acid (HOCl), histidine and tryptophan, were also powerful inhibitors of ActPMN killing. Quenchers of hydroxyl radical (·OH), dimethylsulfoxide and sodium benzoate, had less effect, and required higher concentrations. These data suggest that the enhanced killing of Bd by ActPMN involves one or more oxidative mechanisms, and that there is a prominent role for O , either directly or as a precursor of other active oxygen species, a probable role for HO, and possible roles for O, HOCl, and · OH.

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1988-02-01
2022-01-17
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References

  1. Badwey J A, Karnovsky M L. 1980; Active oxygen species and the functions of phagocytic leukocytes. Annual Review of Biochemistry 49:695–726
    [Google Scholar]
  2. Beers R F, Sizer I W. 1952; A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. Journal of Biological Chemistry 195:133–140
    [Google Scholar]
  3. Brummer E, Stevens D A. 1983; Enhancing effect of murine polymorphonuclear neutrophils (PMN) on the multiplication of Blastomyces dermatitidis in vitro and in vivo . Clinical and Experimental Immunology 54:587–594
    [Google Scholar]
  4. Brummer E, Stevens D A. 1984; Activation of murine polymorphonuclear neutrophils for fungicidal activity with super-natants from antigen-stimulated immune spleen cell cultures. Infection and Immunity 45:447–452
    [Google Scholar]
  5. Brummer E, Sugar A M, Stevens D A. 1984; Immunological activation of polymorphonuclear neutrophils for fungal killing: studies with murine cells and Blastomyces dermatitidis in vitro . Journal of Leukocyte Biology 36:505–520
    [Google Scholar]
  6. Brummer E, Sugar A M, Stevens D A. 1985; Enhanced oxidative burst in immunologically activated but not elicited polymorphonuclear leukocytes correlates with fungicidal activity. Infection and Immunity 49:396–401
    [Google Scholar]
  7. Hasty N, Merkel P B, Radlick P, Kearns D R. 1972; Role of azide in singlet oxygen reactions. Reaction of azide with singlet oxygen. Tetrahedron Letters 1:49–52
    [Google Scholar]
  8. Hodgson E K, Fridovich I. 1974; The production of superoxide radical during the decomposition of potassium peroxochro-mate (V). Biochemistry 13:3811–3815
    [Google Scholar]
  9. Johnston R B. 1975; The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. Journal of Clinical Investigation 55:1357–1372
    [Google Scholar]
  10. Klebanoff S J, Clark R A. 1978 The neutrophil: function and clinical disorders North-Holland, Amsterdam:283
    [Google Scholar]
  11. Lehrer RI, Ladra K M, Hake R B. 1975; Nonoxidative fungicidal mechanisms of mammalian granulocytes : demonstration of components with candidacidal activity in human, rabbit, and guinea pig leukocytes. Infection and Immunity 11:1226–1234
    [Google Scholar]
  12. Lowry O H, Rosebrough N J, Farr A L, Randall R J. 1951; Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  13. Morozumi P A, Brummer E, Stevens D A. 1982; Protection against pulmonary blastomycosis : correlation with cellular and humoral immunity in mice after subcutaneous nonlethal infection. Infection and Immunity 37:670–678
    [Google Scholar]
  14. Sasada M, Johnston R B. 1980; Macrophagemicrobicidal activity : correlation between phagocytosis-associated oxidative metabolism and the killing of Candida by macrophages. Journal of Experimental Medicine 152:85–98
    [Google Scholar]
  15. Singh H, Vadas J A. 1978; Singlet oxygen: a major reactive species in the furocoumarin photosensitized inactivation of E. coli ribosomes. Photochemistry and Photobiology 28:539–545
    [Google Scholar]
  16. Spitznagel J K, Shafer W M. 1985; Neutrophil killing of bacteria by oxygen-independent mechanisms : a historical summary. Reviews of Infectious Diseases 7:398–403
    [Google Scholar]
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