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. 1981 May 1;153(5):1302–1315. doi: 10.1084/jem.153.5.1302

Susceptibility of leishmania to oxygen intermediates and killing by normal macrophages

HW Murray
PMCID: PMC2186145  PMID: 7252418

Abstract

This study demonstrates that the promastigote form of virulent Leishmania donovani and Leishmania tropica are both deficient in endogenous enzymatic scavengers of H(2)0(2) (catalase, glutathione peroxidase) and susceptible to low fluxes of H(2)O(2) in a cell-free model. In addition, the killing of promastigotes by H(2)0(2) is markedly enhanced in the presence of a peroxidase and halide. Promastigotes also readily trigger the macrophage oxidative burst including the generation of H(2)0(2), and most intracellular promastigotes are killed within 18 h by unstimulated normal resident cells. Catalase, but not scavengers or quenchers of O(2)(-), OHx, or (1)O(2), protected promastigotes in a cell-free xanthine oxidase microbicidal system, and catalase also partially inhibited the leishmanicidal activity of resident macrophages. Thus, amongst various oxygen intermediates, H(2)0(2) alone appeared to be both necessary and sufficient for promastigote killing. Depriving macrophages of exogenous glucose, which inhibits the generation of oxygen intermediates, achieved effects similar to catalase treatment. These observations directly contrast with the intracellular parasite, T. gondii which is richly endowed with catalase and glutathione peroxidase, highly resistant to H(2)0(2), and requires products of O(2)(-)-H(2)0(2) interaction for effective oxidative killing. Toxoplasmas also fail to trigger the respiratory burst of normal macrophages, and readily multiply within these cells (1-5). Macrophages first activated by in vivo or in vitro immunologic stimuli, however, display an enhanced capacity to generate oxygen intermediates beyond O(2)(-) and H(2)0(2), and are able to kill toxoplasmas or inhibit their intracellular replication (1, 2). These studies illustrate the wide spectrum of susceptibility to oxidative products which appears to exist for virulent intracellular protozoans, and indicate that such differences may be reflected in contrasting fates of parasites within cell-free oxidative environments and the cytoplasm of normal resident macrophages. In addition, these observations also demonstrate that nonactivated phagocytes may display effective microbicidal activity against certain intracellular pathogens utilizing an oxygen-dependent mechanism.

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Selected References

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  1. Akiyama H. J., Haight R. D. Interaction of Leishmania donovani and hamster peritoneal macrophages. A phase-contrast microscopical study. Am J Trop Med Hyg. 1971 Jul;20(4):539–545. doi: 10.4269/ajtmh.1971.20.539. [DOI] [PubMed] [Google Scholar]
  2. Badwey J. A., Karnovsky M. L. Active oxygen species and the functions of phagocytic leukocytes. Annu Rev Biochem. 1980;49:695–726. doi: 10.1146/annurev.bi.49.070180.003403. [DOI] [PubMed] [Google Scholar]
  3. Baehner R. L., Boxer L. A., Davis J. The biochemical basis of nitroblue tetrazolium reduction in normal human and chronic granulomatous disease polymorphonuclear leukocytes. Blood. 1976 Aug;48(2):309–313. [PubMed] [Google Scholar]
  4. Behin R., Mauel J., Sordat B. Leishmania tropica: pathogenicity and in vitro macrophage function in strains of inbred mice. Exp Parasitol. 1979 Aug;48(1):81–91. doi: 10.1016/0014-4894(79)90057-2. [DOI] [PubMed] [Google Scholar]
  5. Berman J. D., Dwyer D. M., Wyler D. J. Multiplication of Leishmania in human macrophages in vitro. Infect Immun. 1979 Oct;26(1):375–379. doi: 10.1128/iai.26.1.375-379.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boveris A., Martino E., Stoppani A. O. Evaluation of the horseradish peroxidase-scopoletin method for the measurement of hydrogen peroxide formation in biological systems. Anal Biochem. 1977 May 15;80(1):145–158. doi: 10.1016/0003-2697(77)90634-0. [DOI] [PubMed] [Google Scholar]
  7. Bradley D. J. Regulation of Leishmania populations within the host. II. genetic control of acute susceptibility of mice to Leishmania donovani infection. Clin Exp Immunol. 1977 Oct;30(1):130–140. [PMC free article] [PubMed] [Google Scholar]
  8. COHEN G., HOCHSTEIN P. GLUTATHIONE PEROXIDASE: THE PRIMARY AGENT FOR THE ELIMINATION OF HYDROGEN PEROXIDE IN ERYTHROCYTES. Biochemistry. 1963 Nov-Dec;2:1420–1428. doi: 10.1021/bi00906a038. [DOI] [PubMed] [Google Scholar]
  9. Farah F. S., Samra S. A., Nuwayri-Salti N. The role of the macrophage in cutaneous leishmaniasis. Immunology. 1975 Oct;29(4):755–764. [PMC free article] [PubMed] [Google Scholar]
  10. Johnston R. B., Jr, Godzik C. A., Cohn Z. A. Increased superoxide anion production by immunologically activated and chemically elicited macrophages. J Exp Med. 1978 Jul 1;148(1):115–127. doi: 10.1084/jem.148.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Keithly J. S. Infectivity of Leishmania donovani amastigotes and promastigotes for golden hamsters. J Protozool. 1976 May;23(2):244–245. doi: 10.1111/j.1550-7408.1976.tb03763.x. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lewis D. H., Peters W. The resistance of intracellular Leishmania parasites to digestion by lysosomal enzymes. Ann Trop Med Parasitol. 1977 Sep;71(3):295–312. doi: 10.1080/00034983.1977.11687192. [DOI] [PubMed] [Google Scholar]
  14. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  15. Miller H. C., Twohy D. W. Infection of macrophages in culture by leptomonads of Leishmania donovani. J Protozool. 1967 Nov;14(4):781–789. doi: 10.1111/j.1550-7408.1967.tb02078.x. [DOI] [PubMed] [Google Scholar]
  16. Murray H. W., Cohn Z. A. Macrophage oxygen-dependent antimicrobial activity. I. Susceptibility of Toxoplasma gondii to oxygen intermediates. J Exp Med. 1979 Oct 1;150(4):938–949. doi: 10.1084/jem.150.4.938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murray H. W., Cohn Z. A. Macrophage oxygen-dependent antimicrobial activity. III. Enhanced oxidative metabolism as an expression of macrophage activation. J Exp Med. 1980 Dec 1;152(6):1596–1609. doi: 10.1084/jem.152.6.1596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murray H. W., Juangbhanich C. W., Nathan C. F., Cohn Z. A. Macrophage oxygen-dependent antimicrobial activity. II. The role of oxygen intermediates. J Exp Med. 1979 Oct 1;150(4):950–964. doi: 10.1084/jem.150.4.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nathan C. F., Silverstein S. C., Brukner L. H., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. II. Hydrogen peroxide as a mediator of cytotoxicity. J Exp Med. 1979 Jan 1;149(1):100–113. doi: 10.1084/jem.149.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nathan C., Nogueira N., Juangbhanich C., Ellis J., Cohn Z. Activation of macrophages in vivo and in vitro. Correlation between hydrogen peroxide release and killing of Trypanosoma cruzi. J Exp Med. 1979 May 1;149(5):1056–1068. doi: 10.1084/jem.149.5.1056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. North R. J. The concept of the activated macrophage. J Immunol. 1978 Sep;121(3):806–809. [PMC free article] [PubMed] [Google Scholar]
  22. PULVERTAFT R. J., HOYLE G. F. Stages in the life-cycle of Leishmania donovani. Trans R Soc Trop Med Hyg. 1960 Mar;54:191–196. doi: 10.1016/0035-9203(60)90057-2. [DOI] [PubMed] [Google Scholar]
  23. Paglia D. E., Valentine W. N. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 1967 Jul;70(1):158–169. [PubMed] [Google Scholar]
  24. Steinman R. M., Cohn Z. A. The interaction of soluble horseradish peroxidase with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Oct;55(1):186–204. doi: 10.1083/jcb.55.1.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wilson C. B., Tsai V., Remington J. S. Failure to trigger the oxidative metabolic burst by normal macrophages: possible mechanism for survival of intracellular pathogens. J Exp Med. 1980 Feb 1;151(2):328–346. doi: 10.1084/jem.151.2.328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zuckerman A. Current status of the immunology of blood and tissue Protozoa. I. Leishmania. Exp Parasitol. 1975 Dec;38(3):370–400. doi: 10.1016/0014-4894(75)90123-x. [DOI] [PubMed] [Google Scholar]

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