1887

Abstract

The amoeba generated both luminol- and lucigenin-enhanced chemiluminescence upon addition of the respiratory inhibitor sodium cyanide, but not upon the addition of sodium azide. Photon emission in the presence of lucigenin was three- to fourfold greater than that measured in the presence of luminol, but both forms of chemiluminescence were strictly dependent upon the presence of O, indicating the requirement for oxidative reactions in these processes. Lucigenin-chemilununescence measured during the phagocytosis of latex beads under identical conditions was, however, barely detectable above background levels. Cyanide similarly induced the formation of , as indicated by the stimulation of superoxide-dismutase-inhibitable cytochrome reduction, and the rate of production was similar to that previously observed during phagocytosis of latex particles or yeasts by these cells. Thus, in view of the similar rates of production during cyanide exposure or phagocytosis, but disparate rates of lucigenin-chemiluminescence, these two treatments must activate different molecular processes leading to reactive oxidant production. The rates of cyanide-stimulated lucigenin-chemiluminescence were directly proportional to the O tensions in the medium from 0 to 300 μM, indicating that the rates of free-radical-generating reactions were directly related to the oxygen tensions in the environment. The superoxide dismutase inhibitor diethyldithiocarbamate similarly stimulated lucigenin-chemiluminescence, with photon emission again being dependent upon O tensions in the range 0–320 μM. A mechanism by which cells may limit generating reactions, and so reduce damaging free radical reactions, was observed when anaerobic suspensions were reaerated. These data indicate that oxidative stress and phagocytosis provide two intracellular sources of free-radical generating reactions in these cells.

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/content/journal/micro/10.1099/00221287-137-5-1021
1991-05-01
2021-04-23
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References

  1. Allen R. C. 1981; Lucigenin chemiluminescence: a new approach to the study of polymorphonuclear leukocyte redox activity. In Bioluminescence and Chemiluminescence63–73 De Luca M. A. New York: Academic Press;
    [Google Scholar]
  2. Allen R. C., Loose L. D. 1976; Phagocytic activation of a luminol-dependent chemiluminescence in rabbit alveolar and peritoneal macrophages. Biochimica et Biophysica Acta 96:245–252
    [Google Scholar]
  3. Babior B. M., Kipnes R. S., Curnutte J. T. 1973; Biological defense mechanisms: the production by leukocytes of superoxide, a potential bactericidal agent. Journal of Clinical Investigation 52:741–744
    [Google Scholar]
  4. Curnutte J. T. 1988; Phagocyte Defects I and II. Hematology I Oncology Clinics of North America Philadelphia, USA: Saunders;
    [Google Scholar]
  5. Davies B., Chattings L. S., Edwards S. W. 1991; Superoxide generation during phagocytosis by Acanthamoeba castellanii: similarities to the respiratory burst of immune phagocytes. Journal of General Microbiology 137:705–710
    [Google Scholar]
  6. Edwards S. W. 1987; Luminol- and lucigenin-dependent chemiluminescence of neutrophils: role of degranulation. Journal of Clinical and Laboratory Immunology 22:35–39
    [Google Scholar]
  7. Edwards S. W., Lloyd D. 1977; Cyanide-insensitive respiration in Acanthamoeba castellanii. Changes in sensitivity of whole cell respiration during exponential growth. Journal of General Microbiology 103:207–213
    [Google Scholar]
  8. Edwards S. W., Lloyd D. 1978; Properties of mitochondria isolated from cyanide-sensitive and cyanide-stimulated cultures of Acanthamoeba castellanii. Biochemical Journal 174:203–211
    [Google Scholar]
  9. Edwards S. W., Say J. E., Hart C. A. 1987; Oxygen-dependent killing of Staphylococcus aureus by human neutrophils. Journal of General Microbiology 133:3591–3597
    [Google Scholar]
  10. Halliwell B., Gutteridge J. M. C. 1985 Free Radicals in Biology and Medicine Oxford: Clarendon Press;
    [Google Scholar]
  11. Lloyd D., Edwards S. W., Kristensen B., Degn H. 1979a; The effects of inhibitors on the oxygen kinetics of terminal oxidases of Acanthamoeba castellanii. Biochemical Journal 182:11–15
    [Google Scholar]
  12. Lloyd D., Boveris A., Reiter R., Filipowski M., Chance B. 1979b; Chemiluminescence in Acanthamoeba castellanii. Biochemical Journal 184:149–156
    [Google Scholar]
  13. Lloyd D., Mellor H., Williams J. L. 1983a; Oxygen affinity of the respiratory chain of Acanthamoeba castellanii. Biochemical Journal 214:47–51
    [Google Scholar]
  14. Lloyd D., Protheroe R., Williams T. N., Williams J. L. 1983b; Adaptation of the respiratory system of Acanthamoeba castellanii to anaerobiosis. FEMS Microbiology Letters 17:143–146
    [Google Scholar]
  15. Segal A. W., Jones O. T. G. 1978 Novel cytochrome b system in phagocytic vesicles of human granulocytes Nature; London: 276:515–517
    [Google Scholar]
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