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A study of the sensitivity of Leishmania donovani promastigotes and amastigotes to hydrogen peroxide. I. Differences in sensitivity correlate with parasite-mediated removal of hydrogen peroxide

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

The sensitivities of promastigotes and amastigotes of Leishmania donovani to reagent or glucose oxidase-generated hydrogen peroxide (H2O2) were examined in a phagocyte-free system and compared with direct measurements of loss of H2O2 due to reaction with the parasite. Using a combined fluorescence dye uptake/dye exclusion viability assay in conjunction with motility and transformation data it was shown that log-phase promastigotes harvested from recently transformed cultures were intermediate in their H2O2 sensitivity between amastigotes and log-phase promastigotes harvested from long-term subcultures. It was also observed that, while promastigotes are equally sensitive to either form of H2O2 stress, amastigotes are more resistant to single larger amounts of reagent H2O2 than to equivalent amounts of H2O2 generated over a 1 h period. In each case the respective LD50 values obtained for each form of the parasite under each type of H2O2 stress correlated with saturation of their ability to remove H2O2 from the phagocyte-free system. For both promastigotes and amastigotes there was always a time delay after removal of either form of H2O2 stress before H2O2-mediated damage to membranes became apparent. The results suggest that the differential responses of promastigotes and amastigotes to different forms of H2O2 stress may depend upon different H2O2 scavenging mechanisms examined in more detail in the accompanying paper.

Type
Research Article
Information
Parasitology , Volume 91 , Issue 2 , October 1985 , pp. 197 - 206
Copyright
Copyright © Cambridge University Press 1985

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References

REFERENCES

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
Burchill, B. R., Oliver, J. M., Pearson, C. B., Leinbach, E. D. & Berlin, R. D. (1978). Microtubule dynamics and glutathione metabolism in phagocytozing human polymorphonuclear leucocytes. Journal of Cell Biology 76, 439–47.CrossRefGoogle Scholar
Channon, J. Y., Roberts, M. V. & Blackwell, J. M. (1984). A study of the differential respiratory burst activity elicited by promastigotes and amastigotes of Leishmania donovani in murine resident peritoneal macrophages. Immunology 53, 345–55.Google ScholarPubMed
Cochbane, C. G., Bragg, R. G., Fehraufstatta, I. U., Revak, S. D., Hyslop, P. A. & Hinshaw, D. B. (1985). Oxidant and protease effectors in acute inflammation. In Proceedings of the Fourth Leiden Conference on Mononuclear Phagocytes (ed. Van Furth, R.), The Hague, Martinus Nijhoff Publishers BV.Google Scholar
Doran, T. I. & Herman, R. (1981). Characterization of populations of promastigotes of Leishmania donovani. Journal of Protozoology 28, 345–50.CrossRefGoogle ScholarPubMed
Giannini, M. S. (1974). Effects of promastigote growth phase, frequency of subculture, and host age on promastigote–initiated infections with Leishmania donovani in the golden hamster. Journal of Protozoology 21, 521–7.CrossRefGoogle ScholarPubMed
Gutteridge, J. M. C. & Wilkins, S. (1983). Copper salt-dependent hydroxyl radical formation damage to proteins acting as antioxidants. Biochimica et Biophysica acta 759, 38–41.CrossRefGoogle ScholarPubMed
Hart, D. T., Vickerman, K. & Coombs, G. H. (1981). Transformation in vitro of Leishmania mexicana amastigotes to promastigotes: nutritional requirements and the effects of drugs. Parasitology 83, 529–41.CrossRefGoogle Scholar
Johnston, R. B., Godzik, C. A. & Cohn, Z. A. (1978). Increased superoxide anion production by immunologically activated and chemically elicited macrophages. Journal of Experimental Medicine 148, 115–27.CrossRefGoogle ScholarPubMed
Kosoweb, N. S. & Kosoweb, E. M. (1978). The glutathione status of cells. International Review of Cytology 54, 109–60.CrossRefGoogle Scholar
Maehly, A. C. & Chance, B. (1954). Catalases and peroxidases. Methods of Biochemical Analysis 1, 357–424.CrossRefGoogle ScholarPubMed
McCord, J. M. & Day, E. D. (1978). Superoxide-dependent production of hydroxyl radical catalyzed by iron-EDTA complex. FEBS Letters 86, 139–42.CrossRefGoogle ScholarPubMed
Mubbay, H. W. (1981). Susceptibility of Leishmania to oxygen intermediates and killing by normal macrophages. Journal of Experimental Medicine 153, 1302–15.Google Scholar
Mubbay, 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.Google Scholar
Nathan, C. F. & Root, R. K. (1977). Hydrogen peroxide release from mouse peritoneal macrophages. Dependence on sequential activation and triggering. Journal of Experimental Medicine 146, 1648–62.CrossRefGoogle ScholarPubMed
Peabson, 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 Scholar
Reineb, N. E. & Kazuba, J. W. (1982). Oxidant-mediated damage of Leishmania donovani promastigotes. Infection and Immunity 36, 1023–7.Google Scholar
Sacks, D. L. & Perkins, P. V. (1984). Identification of an infective stage of Leishmania promastigotes. Science 223, 1417–19.CrossRefGoogle ScholarPubMed
Steiger, R. F. & Black, C. D. V. (1980). Simplified defined media for cultivating Leishmania donovani promastigotes. Acta, Tropica 37, 195–8.Google ScholarPubMed
Tappel, A. L. (1973). Lipid peroxidation damage to cell components. Federation Proceedings 32, 1870–4.Google ScholarPubMed