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. 1991 Feb;59(2):521–528. doi: 10.1128/iai.59.2.521-528.1991

Mechanism for candidacidal activity in macrophages activated by recombinant gamma interferon.

K Watanabe 1, K Kagaya 1, T Yamada 1, Y Fukazawa 1
PMCID: PMC257780  PMID: 1898907

Abstract

Candidacidal activity in macrophages activated by recombinant gamma interferon was examined kinetically in relation to acidification of phagolysosomes. In resident peritoneal macrophages (PMPs) of BALB/c mice, enhanced killing activity against Candida albicans was demonstrated after incubation with 100 U of gamma interferon per ml for 24 h but not after incubation for 48 to 72 h. Conversely, increased generation of H2O2 was exhibited in PMPs incubated from 48 to 72 h but not in PMPs incubated for 24 h. In normal PMPs, fusion of lysosomes to candida-containing phagosomes was readily accomplished and phagosome-lysosome fusion was not enhanced further by activation. The candidacidal substance was extracted from granule-rich fractions of either normal or activated PMPs by using citric acid (pH 2.7) in equal amounts; the substance showed a noncationic, heat-stable protein nature. In addition, when phagolysosomal pH was determined by flow cytometry of intraphagolysosomal fluorescein isothiocyanate-labeled C. albicans, phagolysosomes with low pH (less than 4.0) were detected in about 40% of PMPs activated for 24 h but not in those activated for 72 h or in normal PMPs. Moreover, increasing the intralysosomal pH with NH4Cl resulted in a significant reduction of candidacidal activity in activated PMPs. These results indicate that the candidacidal activity of gamma interferon-activated PMPs correlates well with enhanced acidification of their phagolysosomes and suggest that the candidacidal activity of activated PMPs is independent from reactive oxygen molecules and is mediated by proteinaceous substance(s) generated only in a strong acidic milieu of phagolysosomes by activation.

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Selected References

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  1. Arai T., Mikami Y., Yokoyama K. Phagocytosis of Candida albicans by rabbit alveolar macrophages and guinea pig neutrophils. Sabouraudia. 1977 Jul;15(2):171–177. [PubMed] [Google Scholar]
  2. Araki S., Kagaya K., Kitoh K., Kimura M., Fukazawa Y. Enhancement of resistance to Escherichia coli infection in mice by dihydroheptaprenol, a synthetic polyprenol derivative. Infect Immun. 1987 Sep;55(9):2164–2170. doi: 10.1128/iai.55.9.2164-2170.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Armstrong J. A., Hart P. D. Response of cultured macrophages to Mycobacterium tuberculosis, with observations on fusion of lysosomes with phagosomes. J Exp Med. 1971 Sep 1;134(3 Pt 1):713–740. doi: 10.1084/jem.134.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baccarini M., Blasi E., Puccetti P., Bistoni F. Phagocytic killing of Candida albicans by different murine effector cells. Sabouraudia. 1983 Dec;21(4):271–286. [PubMed] [Google Scholar]
  5. Baghian A., Lee K. W. Role of activated macrophages in resistance to systemic candidosis. J Leukoc Biol. 1988 Sep;44(3):166–171. doi: 10.1002/jlb.44.3.166. [DOI] [PubMed] [Google Scholar]
  6. Beaman L., Benjamini E., Pappagianis D. Activation of macrophages by lymphokines: enhancement of phagosome-lysosome fusion and killing of Coccidioides immitis. Infect Immun. 1983 Mar;39(3):1201–1207. doi: 10.1128/iai.39.3.1201-1207.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bhisey A. N., Freed J. J. Altered movement of endosomes in colchicine-treated cultured macrophages. Exp Cell Res. 1971 Feb;64(2):430–438. doi: 10.1016/0014-4827(71)90097-8. [DOI] [PubMed] [Google Scholar]
  8. Brummer E., Morrison C. J., Stevens D. A. Recombinant and natural gamma-interferon activation of macrophages in vitro: different dose requirements for induction of killing activity against phagocytizable and nonphagocytizable fungi. Infect Immun. 1985 Sep;49(3):724–730. doi: 10.1128/iai.49.3.724-730.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brummer E., Stevens D. A. Candidacidal mechanisms of peritoneal macrophages activated with lymphokines or gamma-interferon. J Med Microbiol. 1989 Mar;28(3):173–181. doi: 10.1099/00222615-28-3-173. [DOI] [PubMed] [Google Scholar]
  10. Collart M. A., Belin D., Vassalli J. D., de Kossodo S., Vassalli P. Gamma interferon enhances macrophage transcription of the tumor necrosis factor/cachectin, interleukin 1, and urokinase genes, which are controlled by short-lived repressors. J Exp Med. 1986 Dec 1;164(6):2113–2118. doi: 10.1084/jem.164.6.2113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Diamond R. D., Huber E., Haudenschild C. C. Mechanisms of destruction of Aspergillus fumigatus hyphae mediated by human monocytes. J Infect Dis. 1983 Mar;147(3):474–483. doi: 10.1093/infdis/147.3.474. [DOI] [PubMed] [Google Scholar]
  12. Farahbakhsh Z. T., Baldwin R. L., Wisnieski B. J. Effect of low pH on the conformation of Pseudomonas exotoxin A. J Biol Chem. 1987 Feb 15;262(5):2256–2261. [PubMed] [Google Scholar]
  13. Flesch I. E., Kaufmann S. H. Attempts to characterize the mechanisms involved in mycobacterial growth inhibition by gamma-interferon-activated bone marrow macrophages. Infect Immun. 1988 Jun;56(6):1464–1469. doi: 10.1128/iai.56.6.1464-1469.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flesch I. E., Schwamberger G., Kaufmann S. H. Fungicidal activity of IFN-gamma-activated macrophages. Extracellular killing of Cryptococcus neoformans. J Immunol. 1989 May 1;142(9):3219–3224. [PubMed] [Google Scholar]
  15. Fukazawa Y., Kagaya K. Host defence mechanisms against fungal infection. Microbiol Sci. 1988 Apr;5(4):124–127. [PubMed] [Google Scholar]
  16. Giger D. K., Domer J. E., Moser S. A., McQuitty J. T., Jr Experimental murine candidiasis: pathological and immune responses in T-lymphocyte-depleted mice. Infect Immun. 1978 Sep;21(3):729–737. doi: 10.1128/iai.21.3.729-737.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gray P. W., Goeddel D. V. Cloning and expression of murine immune interferon cDNA. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5842–5846. doi: 10.1073/pnas.80.19.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Horwitz M. A. The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes. J Exp Med. 1983 Dec 1;158(6):2108–2126. doi: 10.1084/jem.158.6.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jacques Y. V., Bainton D. F. Changes in pH within the phagocytic vacuoles of human neutrophils and monocytes. Lab Invest. 1978 Sep;39(3):179–185. [PubMed] [Google Scholar]
  20. Kagaya K., Shinoda T., Fukazawa Y. Murine defense mechanism against Candida albicans infection. I. Collaboration of cell-mediated and humoral immunities in protection against systemic C. albicans infection. Microbiol Immunol. 1981;25(7):647–654. doi: 10.1111/j.1348-0421.1981.tb00068.x. [DOI] [PubMed] [Google Scholar]
  21. Kagaya K., Watanabe K., Fukazawa Y. Capacity of recombinant gamma interferon to activate macrophages for Salmonella-killing activity. Infect Immun. 1989 Feb;57(2):609–615. doi: 10.1128/iai.57.2.609-615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Karbassi A., Becker J. M., Foster J. S., Moore R. N. Enhanced killing of Candida albicans by murine macrophages treated with macrophage colony-stimulating factor: evidence for augmented expression of mannose receptors. J Immunol. 1987 Jul 15;139(2):417–421. [PubMed] [Google Scholar]
  23. 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]
  24. Lehrer R. I., Ganz T., Szklarek D., Selsted M. E. Modulation of the in vitro candidacidal activity of human neutrophil defensins by target cell metabolism and divalent cations. J Clin Invest. 1988 Jun;81(6):1829–1835. doi: 10.1172/JCI113527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lehrer R. I., Ladra K. M., Hake R. B. Nonoxidative fungicidal mechanisms of mammalian granulocytes: demonstration of components with candidacidal activity in human, rabbit, and guinea pig leukocytes. Infect Immun. 1975 Jun;11(6):1226–1234. doi: 10.1128/iai.11.6.1226-1234.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lehrer R. I., Szklarek D., Selsted M. E., Fleischmann J. Increased content of microbicidal cationic peptides in rabbit alveolar macrophages elicited by complete Freund adjuvant. Infect Immun. 1981 Sep;33(3):775–778. doi: 10.1128/iai.33.3.775-778.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mor N., Goren M. B. Discrepancy in assessment of phagosome-lysosome fusion with two lysosomal markers in murine macrophages infected with Candida albicans. Infect Immun. 1987 Jul;55(7):1663–1667. doi: 10.1128/iai.55.7.1663-1667.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nathan C. F., Murray H. W., Wiebe M. E., Rubin B. Y. Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J Exp Med. 1983 Sep 1;158(3):670–689. doi: 10.1084/jem.158.3.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ohkuma S., Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3327–3331. doi: 10.1073/pnas.75.7.3327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pace J. L., Russell S. W., Torres B. A., Johnson H. M., Gray P. W. Recombinant mouse gamma interferon induces the priming step in macrophage activation for tumor cell killing. J Immunol. 1983 May;130(5):2011–2013. [PubMed] [Google Scholar]
  31. Patterson-Delafield J., Martinez R. J., Lehrer R. I. Microbicidal cationic proteins in rabbit alveolar macrophages: a potential host defense mechanism. Infect Immun. 1980 Oct;30(1):180–192. doi: 10.1128/iai.30.1.180-192.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Perfect J. R., Granger D. L., Durack D. T. Effects of antifungal agents and gamma interferon on macrophage cytotoxicity for fungi and tumor cells. J Infect Dis. 1987 Aug;156(2):316–323. doi: 10.1093/infdis/156.2.316. [DOI] [PubMed] [Google Scholar]
  33. Pick E., Keisari Y. A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture. J Immunol Methods. 1980;38(1-2):161–170. doi: 10.1016/0022-1759(80)90340-3. [DOI] [PubMed] [Google Scholar]
  34. Poole B., Ohkuma S. Effect of weak bases on the intralysosomal pH in mouse peritoneal macrophages. J Cell Biol. 1981 Sep;90(3):665–669. doi: 10.1083/jcb.90.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sasada M., Kubo A., Nishimura T., Kakita T., Moriguchi T., Yamamoto K., Uchino H. Candidacidal activity of monocyte-derived human macrophages: relationship between Candida killing and oxygen radical generation by human macrophages. J Leukoc Biol. 1987 Apr;41(4):289–294. doi: 10.1002/jlb.41.4.289. [DOI] [PubMed] [Google Scholar]

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