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. 1995 May;63(5):2075–2078. doi: 10.1128/iai.63.5.2075-2078.1995

Lack of involvement of nitric oxide in killing of Aspergillus fumigatus conidia by pulmonary alveolar macrophages.

E Michaliszyn 1, S Sénéchal 1, P Martel 1, L de Repentigny 1
PMCID: PMC173267  PMID: 7729922

Abstract

Nitric oxide is an important antimicrobial mechanism of phagocytes from mice and rats, but in the case of human phagocytes, secretion is still controversial. We investigated whether nitric oxide is involved in the killing of Aspergillus fumigatus conidia by human or murine pulmonary alveolar macrophages. Stimulation of the macrophages with gamma interferon and Escherichia coli lipopolysaccharide had no effect on fungicidal activity against conidia in vitro, with or without the addition of tetrahydrobiopterin. Killing of conidia (means +/- standard deviations) by murine or human alveolar macrophages, before and after stimulation, was 44% +/- 13% and 49% +/- 12% (P = 0.34) and 24% +/- 5% and 29% +/- 10% (P = 0.20), respectively. Fungicidal activity was unaltered in the presence of the competitive inhibitor NG-monomethyl L-arginine, and nitrite was undetectable in cell supernatants. Peritoneal macrophages from B6C3F1 mice produced 18 mumol of nitrite per 10(6) cells in 18 h. In conclusion, nitric oxide does not appear to be involved in the fungicidal activity of murine or human alveolar macrophages against A. fumigatus conidia.

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Selected References

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  1. Alspaugh J. A., Granger D. L. Inhibition of Cryptococcus neoformans replication by nitrogen oxides supports the role of these molecules as effectors of macrophage-mediated cytostasis. Infect Immun. 1991 Jul;59(7):2291–2296. doi: 10.1128/iai.59.7.2291-2296.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cameron M. L., Granger D. L., Weinberg J. B., Kozumbo W. J., Koren H. S. Human alveolar and peritoneal macrophages mediate fungistasis independently of L-arginine oxidation to nitrite or nitrate. Am Rev Respir Dis. 1990 Dec;142(6 Pt 1):1313–1319. doi: 10.1164/ajrccm/142.6_Pt_1.1313. [DOI] [PubMed] [Google Scholar]
  3. Denis M. Tumor necrosis factor and granulocyte macrophage-colony stimulating factor stimulate human macrophages to restrict growth of virulent Mycobacterium avium and to kill avirulent M. avium: killing effector mechanism depends on the generation of reactive nitrogen intermediates. J Leukoc Biol. 1991 Apr;49(4):380–387. doi: 10.1002/jlb.49.4.380. [DOI] [PubMed] [Google Scholar]
  4. Di Rosa M., Radomski M., Carnuccio R., Moncada S. Glucocorticoids inhibit the induction of nitric oxide synthase in macrophages. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1246–1252. doi: 10.1016/0006-291x(90)91583-e. [DOI] [PubMed] [Google Scholar]
  5. Fischer-Stenger K., Marciano-Cabral F. The arginine-dependent cytolytic mechanism plays a role in destruction of Naegleria fowleri amoebae by activated macrophages. Infect Immun. 1992 Dec;60(12):5126–5131. doi: 10.1128/iai.60.12.5126-5131.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fortier A. H., Polsinelli T., Green S. J., Nacy C. A. Activation of macrophages for destruction of Francisella tularensis: identification of cytokines, effector cells, and effector molecules. Infect Immun. 1992 Mar;60(3):817–825. doi: 10.1128/iai.60.3.817-825.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fuchs D., Murr C., Reibnegger G., Weiss G., Werner E. R., Werner-Felmayer G., Wachter H. Nitric oxide synthase and antimicrobial armature of human macrophages. J Infect Dis. 1994 Jan;169(1):224–225. doi: 10.1093/infdis/169.1.224. [DOI] [PubMed] [Google Scholar]
  8. Granger D. L., Hibbs J. B., Jr, Perfect J. R., Durack D. T. Metabolic fate of L-arginine in relation to microbiostatic capability of murine macrophages. J Clin Invest. 1990 Jan;85(1):264–273. doi: 10.1172/JCI114422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Granger D. L., Hibbs J. B., Jr, Perfect J. R., Durack D. T. Specific amino acid (L-arginine) requirement for the microbiostatic activity of murine macrophages. J Clin Invest. 1988 Apr;81(4):1129–1136. doi: 10.1172/JCI113427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Green L. C., Wagner D. A., Glogowski J., Skipper P. L., Wishnok J. S., Tannenbaum S. R. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem. 1982 Oct;126(1):131–138. doi: 10.1016/0003-2697(82)90118-x. [DOI] [PubMed] [Google Scholar]
  11. Green S. J., Meltzer M. S., Hibbs J. B., Jr, Nacy C. A. Activated macrophages destroy intracellular Leishmania major amastigotes by an L-arginine-dependent killing mechanism. J Immunol. 1990 Jan 1;144(1):278–283. [PubMed] [Google Scholar]
  12. Hibbs J. B., Jr, Taintor R. R., Vavrin Z. Macrophage cytotoxicity: role for L-arginine deiminase and imino nitrogen oxidation to nitrite. Science. 1987 Jan 23;235(4787):473–476. doi: 10.1126/science.2432665. [DOI] [PubMed] [Google Scholar]
  13. Hibbs J. B., Jr, Taintor R. R., Vavrin Z., Rachlin E. M. Nitric oxide: a cytotoxic activated macrophage effector molecule. Biochem Biophys Res Commun. 1988 Nov 30;157(1):87–94. doi: 10.1016/s0006-291x(88)80015-9. [DOI] [PubMed] [Google Scholar]
  14. Iyengar R., Stuehr D. J., Marletta M. A. Macrophage synthesis of nitrite, nitrate, and N-nitrosamines: precursors and role of the respiratory burst. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6369–6373. doi: 10.1073/pnas.84.18.6369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jorens P. G., Van Overveld F. J., Bult H., Vermeire P. A., Herman A. G. L-arginine-dependent production of nitrogen oxides by rat pulmonary macrophages. Eur J Pharmacol. 1991 Aug 6;200(2-3):205–209. doi: 10.1016/0014-2999(91)90573-9. [DOI] [PubMed] [Google Scholar]
  16. Karupiah G., Xie Q. W., Buller R. M., Nathan C., Duarte C., MacMicking J. D. Inhibition of viral replication by interferon-gamma-induced nitric oxide synthase. Science. 1993 Sep 10;261(5127):1445–1448. doi: 10.1126/science.7690156. [DOI] [PubMed] [Google Scholar]
  17. Kawabe T., Isobe K. I., Hasegawa Y., Nakashima I., Shimokata K. Immunosuppressive activity induced by nitric oxide in culture supernatant of activated rat alveolar macrophages. Immunology. 1992 May;76(1):72–78. [PMC free article] [PubMed] [Google Scholar]
  18. Levitz S. M., Selsted M. E., Ganz T., Lehrer R. I., Diamond R. D. In vitro killing of spores and hyphae of Aspergillus fumigatus and Rhizopus oryzae by rabbit neutrophil cationic peptides and bronchoalveolar macrophages. J Infect Dis. 1986 Sep;154(3):483–489. doi: 10.1093/infdis/154.3.483. [DOI] [PubMed] [Google Scholar]
  19. Li Y., Severn A., Rogers M. V., Palmer R. M., Moncada S., Liew F. Y. Catalase inhibits nitric oxide synthesis and the killing of intracellular Leishmania major in murine macrophages. Eur J Immunol. 1992 Feb;22(2):441–446. doi: 10.1002/eji.1830220223. [DOI] [PubMed] [Google Scholar]
  20. Liew F. Y., Cox F. E. Nonspecific defence mechanism: the role of nitric oxide. Immunol Today. 1991 Mar;12(3):A17–A21. doi: 10.1016/S0167-5699(05)80006-4. [DOI] [PubMed] [Google Scholar]
  21. Liew F. Y., Li Y., Moss D., Parkinson C., Rogers M. V., Moncada S. Resistance to Leishmania major infection correlates with the induction of nitric oxide synthase in murine macrophages. Eur J Immunol. 1991 Dec;21(12):3009–3014. doi: 10.1002/eji.1830211216. [DOI] [PubMed] [Google Scholar]
  22. Liew F. Y., Millott S., Parkinson C., Palmer R. M., Moncada S. Macrophage killing of Leishmania parasite in vivo is mediated by nitric oxide from L-arginine. J Immunol. 1990 Jun 15;144(12):4794–4797. [PubMed] [Google Scholar]
  23. Lorsbach R. B., Russell S. W. A specific sequence of stimulation is required to induce synthesis of the antimicrobial molecule nitric oxide by mouse macrophages. Infect Immun. 1992 May;60(5):2133–2135. doi: 10.1128/iai.60.5.2133-2135.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lowenstein C. J., Dinerman J. L., Snyder S. H. Nitric oxide: a physiologic messenger. Ann Intern Med. 1994 Feb 1;120(3):227–237. doi: 10.7326/0003-4819-120-3-199402010-00009. [DOI] [PubMed] [Google Scholar]
  25. Marletta M. A. Nitric oxide synthase structure and mechanism. J Biol Chem. 1993 Jun 15;268(17):12231–12234. [PubMed] [Google Scholar]
  26. Martin J. H., Edwards S. W. Changes in mechanisms of monocyte/macrophage-mediated cytotoxicity during culture. Reactive oxygen intermediates are involved in monocyte-mediated cytotoxicity, whereas reactive nitrogen intermediates are employed by macrophages in tumor cell killing. J Immunol. 1993 Apr 15;150(8 Pt 1):3478–3486. [PubMed] [Google Scholar]
  27. Moncada S., Higgs A. The L-arginine-nitric oxide pathway. N Engl J Med. 1993 Dec 30;329(27):2002–2012. doi: 10.1056/NEJM199312303292706. [DOI] [PubMed] [Google Scholar]
  28. Murray H. W., Teitelbaum R. F. L-arginine-dependent reactive nitrogen intermediates and the antimicrobial effect of activated human mononuclear phagocytes. J Infect Dis. 1992 Mar;165(3):513–517. doi: 10.1093/infdis/165.3.513. [DOI] [PubMed] [Google Scholar]
  29. Olken N. M., Rusche K. M., Richards M. K., Marletta M. A. Inactivation of macrophage nitric oxide synthase activity by NG-methyl-L-arginine. Biochem Biophys Res Commun. 1991 Jun 14;177(2):828–833. doi: 10.1016/0006-291x(91)91864-9. [DOI] [PubMed] [Google Scholar]
  30. Salvemini D., de Nucci G., Gryglewski R. J., Vane J. R. Human neutrophils and mononuclear cells inhibit platelet aggregation by releasing a nitric oxide-like factor. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6328–6332. doi: 10.1073/pnas.86.16.6328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schaffner A., Douglas H., Braude A. Selective protection against conidia by mononuclear and against mycelia by polymorphonuclear phagocytes in resistance to Aspergillus. Observations on these two lines of defense in vivo and in vitro with human and mouse phagocytes. J Clin Invest. 1982 Mar;69(3):617–631. doi: 10.1172/JCI110489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schaffner A. Therapeutic concentrations of glucocorticoids suppress the antimicrobial activity of human macrophages without impairing their responsiveness to gamma interferon. J Clin Invest. 1985 Nov;76(5):1755–1764. doi: 10.1172/JCI112166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schneemann M., Schoedon G., Hofer S., Blau N., Guerrero L., Schaffner A. Nitric oxide synthase is not a constituent of the antimicrobial armature of human mononuclear phagocytes. J Infect Dis. 1993 Jun;167(6):1358–1363. doi: 10.1093/infdis/167.6.1358. [DOI] [PubMed] [Google Scholar]
  34. Sherman M. P., Loro M. L., Wong V. Z., Tashkin D. P. Cytokine- and Pneumocystis carinii- induced L-arginine oxidation by murine and human pulmonary alveolar macrophages. J Protozool. 1991 Nov-Dec;38(6):234S–236S. [PubMed] [Google Scholar]
  35. Stuehr D. J., Marletta M. A. Induction of nitrite/nitrate synthesis in murine macrophages by BCG infection, lymphokines, or interferon-gamma. J Immunol. 1987 Jul 15;139(2):518–525. [PubMed] [Google Scholar]
  36. Stuehr D. J., Marletta M. A. Mammalian nitrate biosynthesis: mouse macrophages produce nitrite and nitrate in response to Escherichia coli lipopolysaccharide. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7738–7742. doi: 10.1073/pnas.82.22.7738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stuehr D. J., Marletta M. A. Synthesis of nitrite and nitrate in murine macrophage cell lines. Cancer Res. 1987 Nov 1;47(21):5590–5594. [PubMed] [Google Scholar]
  38. Summersgill J. T., Powell L. A., Buster B. L., Miller R. D., Ramirez J. A. Killing of Legionella pneumophila by nitric oxide in gamma-interferon-activated macrophages. J Leukoc Biol. 1992 Dec;52(6):625–629. doi: 10.1002/jlb.52.6.625. [DOI] [PubMed] [Google Scholar]
  39. de Repentigny L., Petitbois S., Boushira M., Michaliszyn E., Sénéchal S., Gendron N., Montplaisir S. Acquired immunity in experimental murine aspergillosis is mediated by macrophages. Infect Immun. 1993 Sep;61(9):3791–3802. doi: 10.1128/iai.61.9.3791-3802.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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