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A study of the sensitivity of Leishmania donovani promastigotes and amastigotes to hydrogen peroxide. II. Possible mechanisms involved in protective H2O2 scavenging

Published online by Cambridge University Press:  06 April 2009

Jacqueline Y. Channon  and
Jenefer M. Blackwell
Jacqueline Y. Channon
Affiliation:
Department of Tropical Hygiene, London School of Hygiene and Tropical Medicine, Keppel St (Gower St), London WC1E 7HT
Jenefer M. Blackwell
Affiliation:
Department of Tropical Hygiene, London School of Hygiene and Tropical Medicine, Keppel St (Gower St), London WC1E 7HT
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Extract

Different hydrogen peroxide (H2O2)-scavenging mechanisms, and the conditions under which they operate, have been examined in promastigotes and amastigotes of Leishmania donovani. For promastigotes, the ability of the parasite to remove H2O2 was completely ablated by sonication whereas for sonicated amastigotes substantial loss of H2O2 from the phagocyte-fre¸e test system still occurred. In direct contrast, the ability of amastigotes, but not promastigotes, to remove H2O2 was markedly inhibited by aminotriazole or sodium azide. This suggested a role for haem-containing enzymes, catalase or peroxidases, as a protective H2O2-scavenging mechanism and was consistent with detection of catalase in amastigotes but not promastigotes using a spectrophotometric assay. Both forms of the parasite did, however, show reduced ability to remove H2O2 at 5–7°C indicating that additional enzymatic scavenging mechanisms may operate. Glutathione peroxidase activity was undetectable in either form of the parasite. The total thiol sink, glutathione (GSH) plus protein thiols, was greater in promastigotes but the ability to regenerate GSH via glutathione reductase was equivalent for promastigotes and amastigotes. Less temperature-dependent non-enzymatic mechanisms (e.g. an unsaturated lipid sink) also appear to contribute to removal of H2O2 by both promastigotes and amastigotes. It seems likely, nevertheless, that the difference in H2O2 sensitivity between the two forms of the parasite relates to the activity of the direct H2O2-scavenging enzyme, catalase, which appears to operate more efficiently against a bolus of reagent H2O2.

Type
Research Article
Information
Parasitology , Volume 91 , Issue 2 , October 1985 , pp. 207 - 217
Copyright
Copyright © Cambridge University Press 1985

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References

REFERENCES

Arrick, B. A., Griffith, O. W. & Cerami, A. (1981). Inhibition of glutathione synthesis as a chemotherapeutic strategy for trypanosomiasis. Journal of Experimental Medicine 153, 720–5.CrossRefGoogle ScholarPubMed
Arrick, B. A., Nathan, C. F., Griffith, O. W. & Cohn, Z. A. (1982). Glutathione depletion sensitizes tumour cells to oxidative cytolysis. Journal of Biological Chemistry 257, 1231–7.CrossRefGoogle ScholarPubMed
Boveris, A., Sies, H., Martino, E. E., Docampo, R., Turrens, J. F. & Stoppani, A. O. M. (1980). Deficient metabolic utilization of hydrogen peroxide in Trypanosoma cruzi. The Biochemical Journal 188, 643–8.CrossRefGoogle ScholarPubMed
Brun, R. & Krassner, S. M. (1976). Quantitative ultrastructural investigations of mitochondrial development in L. donovani during transformation. Journal of Protozoology 23, 493–7.CrossRefGoogle Scholar
Channon, J. Y. & Blackwell, J. M. (1985). A study of the sensitivity of Leishmania donovani promastigotes and amastigotes to hydrogen peroxide. I. Differences in sensitivity correlate withParasite-mediated removal of H2O2. Parasitology 91, 197–206.CrossRefGoogle Scholar
Channon, J. Y., Roberts, M. B. & Blackwell, J. M. (1984). A study of the differential respiratory burst activity elicited by promastigotes and amastigotes of Leishmania donovani in murine resident peritoneal macropohages. Immunology 53, 345–55.Google Scholar
Dvorak, A. M., Dvorak, H. F., Peters, S. P., Shulman, E. S., MacGlashan, D. W., Pyne, K., Harvey, V. S., Galli, S. J. & Lichtenstein, L. M. (1983). Lipid bodies: cytoplasmic organelles important to arachidonate metabolism in macrophages and mast cells. Journal of Immunology 131, 2965–76.CrossRefGoogle ScholarPubMed
Griffith, O. W. (1980). Determination of glutathione and glutathione disulphide using glutathione reductase and 2-vinylpyridine. Analytical Biochemistry 106, 207–12.CrossRefGoogle ScholarPubMed
Jackett, P. S., Aber, V. R. & Lowrie, D. B. (1978). Virulence and resistance to superoxide, low ph and hydrogen peroxide among strains of Mycobacterium tuberculosis. Journal of General Microbiology 104, 37–45.CrossRefGoogle Scholar
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, 265–75.CrossRefGoogle ScholarPubMed
Murray, H. W. (1981). Susceptibility of Leishmania to oxygen intermediates and killing by normal macrophages. Journal of Experimental Medicine 153, 1302–15.CrossRefGoogle ScholarPubMed
Murray, H. W. (1982). Cell-mediated immune response in experimental visceral leishmaniasis. II. Oxygen-dependent killing of intracellular Leishmania donovani amastigotes. Journal of Immunology 129, 351–7.CrossRefGoogle ScholarPubMed
Murray, H. W. & Cohn, Z. A. (1979). Macrophage oxygen-dependent antimicrobial activity. 1. Susceptibility of T. gondii to oxygen intermediates. Journal of Experimental Medicine 150, 938–49.CrossRefGoogle ScholarPubMed
Nathan, C. F., Arrick, B. A., Murray, H. W., DeSantis, N. M. & Cohn, Z. A. (1981). Tumour cell anti-oxidant defenses. Inhibition of the glutathione redox cycle enhances macrophage-mediated cytolysis. Journal of Experimental Medicine 153, 766–82.CrossRefGoogle ScholarPubMed
Nishiki, K., Jamieson, D., Oshino, N. & Chance, B. (1976). Oxygen toxicity in the perfused rat liver And lung under hyperbaric conditions. The Biochemical Journal 160, 343–55.CrossRefGoogle ScholarPubMed
Pearson, R. D., Harcus, J. L., Roberts, D. & Donowitz, G. R. (1983). Differential survival of Leishmania donovani amastigotes in human monocytes. Journal of immunology 131, 1994–9.CrossRefGoogle ScholarPubMed
Rudzinska, M. A., D'alesandro, P. A. & Trager, W. (1964). The fine structure of L. donovani and the role of the kinetoplast in the Leishmania-Leptomonad transformation. Journal of Protozoology 11, 166–91.CrossRefGoogle Scholar
Sasada, M. & Johnston, R. B. (1980). Macrophage microbicidal activity. Correlation between phagocytosis-associated oxidative metabolism and the killing of Candida by macrophages. Journal of Experimental Medicine 152, 85–98.CrossRefGoogle ScholarPubMed