Cytokine induced synthesis of nitric oxide from L-arginine: a cytotoxic mechanism that targets intracellular iron

  • J. B. HibbsJr.
Conference paper
Part of the Key Topics in Brain Research book series (KEYTOPICS)


A cytokine inducible high output nitric oxide synthase was recently identified. It is induced by cytokines in macrophages as well as in nonmacrophage cell types. It is a product of the cell-mediated immune response and probably has multiple functional roles. Current experimental results suggest that cytokine induced synthesis of nitric oxide from L-arginine has a role in the defense of the intracellular environment against intracellular microbes. Nitrosylation of intracellular iron, particularly nonheme iron, appears to be a major biochemical fate of nitric oxide synthesized by the cytokine inducible nitric oxide synthase. This results in inhibition of redox enzymes that have nonheme iron essential for catalytic activity. Because nitric oxide is paramagnetic (has an unpaired electron) it can readily react with dioxygen and the superoxide anion to yield toxic products. As a result, the balance between beneficial and deleterious redox reactions involving nitric oxide must be tightly regulated by currently unidentified mechanisms.


Nitric Oxide Tumor Target Cell Francisella Tularensis Nonheme Iron Aconitase Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adams LB, Hibbs JB Jr, Taintor RR, Krahenbuhl JL (1990) Microbiostatic effect of murine macrophages for Toxoplasma gondii: role of synthesis of inorganic nitrogen oxides from L-arginine. J Immunol 144: 2725–2729PubMedGoogle Scholar
  2. Adams LA, Franzblau AG, Vavrin Z, Hibbs JB Jr, Krahenbuhl JL (1991) L-arginine-dependent macrophage effector functions inhibit metabolic activity of Mycobacterium leprae. J Immunol 147: 1642–1646PubMedGoogle Scholar
  3. Amber IJ, Hibbs JB Jr, Taintor RR, Vavrin Z (1988a) L-arginine dependent effector mechanisms is induced in murine adenocarcinoma cells by culture supernotant from cytotoxic activated macrophages. J Leukoc Biol 43: 187–192Google Scholar
  4. Amber IJ, Hibbs JB Jr, Taintor RR, Vavrin Z (1988b) Cytokines induce an L-arginine-dependent effector system in non-macrophage cells. J Leukoc Biol 44: 58–65Google Scholar
  5. Amber IJ, Hibbs JB Jr., Parker CJ, Johnson BB, Taintor RR, Vavrin Z (1991) Activated macrophage condition medium: identification of the soluble factors inducing cytotoxicity and the L-arginine dependent effector mechanism. J Leukoc Biol 49: 610–620PubMedGoogle Scholar
  6. Bastian NR, Xu S, Shao XL, Shelby J, Hibbs JB Jr (1992) Nitric oxide production in response to allogeneic heart transplant in mice. In: Moncada S, Marietta MA, Hibbs JB Jr, Higgs EA (eds) The biology of nitric oxide, vol 2. Enzymology, biochemistry and immunology. Portland Press, London Chapel Hill, pp 273–276Google Scholar
  7. Beckerman KP, Rogers HW, Corbett JA, Schreiber RD, McDaniel ML, Unanue ER (1993) Release of nitric oxide during the T Cell-independent pathway of macrophage activiation: its role in resistance to Listeria monocytogenes. J Immunol 150: 888–895PubMedGoogle Scholar
  8. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA (1990) Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 87: 1620–1624PubMedCrossRefGoogle Scholar
  9. Chan J, Xing Y, Magliozzo RS, Bloom BR (1992) Killing of virulent Mycobacterium tuberculosisby reactive nitrogen intermediates produced by activated murine macrophages. J Exp Med 175: 1111–1122PubMedCrossRefGoogle Scholar
  10. Curran RD, Billiar TR, Stuehr DJ, Simmons RL (1989) Hepatocytes produce nitrogen oxides from L-arginine in response to inflammatory products of kupffer cells. J Exp Med 170: 1796–1774CrossRefGoogle Scholar
  11. Denis M (1991) Interferon-gamma-treated murine macrophages inhibit growth of tubercle bacilli via the generation of reactive nitrogen intermediates. Cell Immunol 132: 150–157PubMedCrossRefGoogle Scholar
  12. Ding AH, Nathan CF, Stuehr DJ (1988) Release of reactive nitrogen intermediates and reactive oxygen intermediates from muse peritoneal macrophages. J Immunol 141: 2407–2414PubMedGoogle Scholar
  13. Drapier J-C, Hibbs JB Jr (1986) Murine cytotoxic activated macrophages inhibit aconitase in tumor cells. Inhibition involves the iron-sulfur prosthetic group and is reversible. J Clin Invest 78: 790–797PubMedCrossRefGoogle Scholar
  14. Drapier J-C, Hibbs JB Jr (1988) Differentiation of murine macrophages to express nonspecific cytotoxicity for tumor cells results in L-arginine-dependent inhibition of mitochondrial iron-sulfur enzymes in the macrophage effect cells. J Immunol 140: 2829–2838PubMedGoogle Scholar
  15. Drapier J-C, Wietzerbin J, Hibbs JB Jr (1988) Interferon-y and tumor necrosis factor induce the L-arginine-dependent cytotoxic effector mechanism in murine macrophages J Immunol 18: 1587–1592Google Scholar
  16. Drapier J-C, Pellat C, Yann H (1991) Generation of EPR-detectable nitrosyl-iron complexes in tumor target cells cocultured with activated macrophages. J Biol Chem 266: 10162–10167PubMedGoogle Scholar
  17. Fortier AH, Polsinelli T, Green SJ, Nacy CA (1992) Activation of macrophages for destruction of Francisella tularensis: identification of cytokines, effector cells, and effector molecules. Infect Immun 60: 817–825PubMedGoogle Scholar
  18. Gaily JA, Montague PR, Reeke GN, Edelman GM (1990) The NO hypothesis: possible effects of a short-lived, rapidly diffusible signal in the development and function of the nervous system. Proc Natl Acad Sci USA 87: 3547–3551CrossRefGoogle Scholar
  19. Granger DL, Taintor RR, Cook JL, Hibbs JB Jr (1980) Injury of neoplastic cells by murine macrophages leads to inhibition of mitochondrial respiration. J Clin Invest 65: 357–370PubMedCrossRefGoogle Scholar
  20. Granger DI, Lehninger Al (1982) Sites of inhibition of mitochondrial electron transport in macrophage-injured neoplastic cells. J Cell Biol 95: 527–535PubMedCrossRefGoogle Scholar
  21. Granger DL, Hibbs JB Jr, Perfect JR, Durack DT (1990) Metabolic fate of L-arginine in relation to microbiostatic capability of macrophages. J Clin Invest 85: 264–273PubMedCrossRefGoogle Scholar
  22. Green SJ, Meltzer MS, Hibbs JB Jr, Nacy CA (1990) Activated macrophages destroy intracellular Leishmania major amastigotes by an L-arginine dependent killing mechanism. J Immunol 144: 278–283PubMedGoogle Scholar
  23. Halliwell B, Gutteridge MC (1990) Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol 186: 1–85PubMedCrossRefGoogle Scholar
  24. Hibbs JB Jr (1992) Overview of cytotoxic mechanisms and defense of the intracellular environment against microbes. In: Moncada S, Marietta MA, Hibbs JB Jr, Higgs EA (eds) The biology of nitric oxide, vol 2. Enzymology, biochemistry and immunology. Portland Press, London Chapel Hill, pp 201–206Google Scholar
  25. Hibbs JB Jr, Granger DL (1982) Activated macrophage-induced cytostasis and inhibition of aerobic energy metabolism in transformed cells: evaluation of lyric and nonlytic target cell responses. In: Mizuno D, Cohn ZA, Takeya K, Ishida N (eds) Self-defence mechanisms. Role of macrophages. University of Tokyo, Tokyo, pp 319–333Google Scholar
  26. Hibbs JB Jr, Lamber LH Jr, Remington RS (1971) Resistance to murine tumors conferred by chronic infection with intracellular protozoa, Toxoplasma gondiiand Besnoitia jellisoni. J Infect Dis 124: 587–592PubMedCrossRefGoogle Scholar
  27. Hibbs JB Jr, Lambert LH Jr, Remington RS (1972) In vitro nonimmunologic destruction of cells with abnormal growth characteristics by adjuvant activated macrophages. Proc Soc Exp Biol Med 139: 1049–1052PubMedGoogle Scholar
  28. Hibbs JB Jr, Taintor RR, Chapman HA Jr, Weinberg JB (1977) Macrophage tumor killing: influence of the local environment. Science 197: 279–282PubMedCrossRefGoogle Scholar
  29. Hibbs JB Jr, Remington JS, Stewart CC (1980) Modulation of immunity and host resistance by micro-organisms. Pharmacol Ther 8: 37–69CrossRefGoogle Scholar
  30. Hibbs JB Jr, Taintor RR, Vavrin Z (1984) Iron depletion: possible cause of tumor cell cytotoxicity induced by activated macrophages. Biochem Biophys Res Commun 123: 716 723Google Scholar
  31. Hibbs JB Jr, Taintor RR, Vavrin Z (1987a) Macrophage cytotoxicity: role for Larginine deiminase activity and imino nitrogen oxidation to nitrite. Science 235: 473–476CrossRefGoogle Scholar
  32. Hibbs JB Jr, Vavrin Z, Taintor RR (1987b) L-arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells. J Immunol 138: 550–565Google Scholar
  33. Hibbs JB Jr, Taintor RR, Vavrin Z, Rachlin EM (1988) Nitric oxide: a cytotoxic activated macrophage effector molecule. Biochem Biophys Res Commun 157: 87–94 [Erratum Biochem Biophys Res Commun (1989) 158: 624]PubMedCrossRefGoogle Scholar
  34. Hibbs JB Jr, Taintor RR, Vavrin Z, Granger DL, Drapier J-C, Amber IJ, Lancaster JR Jr (1990) Synthesis of nitric oxide from a terminal guanidino nitrogen atom of L-arginine: a molecular mechanism regulating cellular proliferation that targets intracellular iron. In: Nitric oxide from L-arginine: a bioregulatory system. Elsevier, New York, pp 189–223Google Scholar
  35. Hibbs JB Jr, Bastian NR, Taintor RR, Vavrin Z, Granger DL (1992a) Activity of the inducible high output nitric oxide synthase during immunological rejection of allogeneic P815 mastocytoma cells in Swiss-Webster mice. In: Moncada S, Marietta MA, Hibbs JB Jr, Higgs EA (eds) The biology of nitric oxide, vol 2. Enzymology, biochemistry and immunology. Portland Press, London Chapel Hill, pp 237–240Google Scholar
  36. Hibbs JB Jr, Granger DL, Krahenbuhl JL, Adams LB (1992b) Synthesis of nitric oxide from L-arginine: a cytotoxic inducible pathway with antimicrobial activity. In: van Furth R (ed) Mononuclear phagocytes: biology of monocytes and macrophages. Kluwer Academic Publishers, Dordrecht Boston London, pp 279–292Google Scholar
  37. Iyengar R, Stuehr DJ, Marietta MA (1987) Macrophage synthesis of nitrite, nitrate, and N-nitrosamines: precursors and role of the respiratory burst. Proc Nati Acad Sci USA 84: 6369–6373CrossRefGoogle Scholar
  38. Kilbourn RG, Belloni P (1990) Endothelial cell production of nitrogen oxides in response to interferon-y in combination with tumor necrosis factor, interleukin-1, or endotoxin. J Natl Can Inst 82: 772–776CrossRefGoogle Scholar
  39. Kosaka H, Kazuhiko I, Kiyohiro I, Itiro T (1979) Stoichiometry of the reaction of exyhemoglobin with nitrite. Biochim Biophys Acta 581: 184–188PubMedGoogle Scholar
  40. Kwon NS, Stuehr DJ, Nathan CF (1991) Inhibition of tumor cell ribonucleotide reductase by macrophage-derived nitric oxide. J Exp Med 174: 761–768PubMedCrossRefGoogle Scholar
  41. Lancaster JR Jr, Hibbs JB Jr (1990) EPR demonstration of iron-nitrosyl complex formation by cytotoxic activated macrophages. Proc Natl Acad Sci USA 87: 1223–1227PubMedCrossRefGoogle Scholar
  42. Lancaster JR Jr, Langrehr JM, Bergonia HA, Murase N, Starzl TE, Simmons RL, Hoffman RA (1992) Detection of iron-nitrosyl complexes by electron paramagnetic resonance spectroscopy during rejection of vascularized allograft in the rat. In: Moncada S, Marietta MA, Hibbs JB Jr, Higgs EA (eds) The biology of nitric oxide, vol 2. Enzymology, biochemistry and immunology. Portland Press, London Chapel Hill, pp 216–219Google Scholar
  43. Leaf CD, Wishnok JS, Tannenbaum SR (1990) In: Moncada S, Higgs EA (eds) Nitric oxide form L-arginine: a bioregulatory system. Elsevier, Amsterdam, pp 291–299Google Scholar
  44. Lepoivre M, Chenais B, Yapo A, Lemaire G, Thelander L, Tenu J-P (1990) Alterations of ribonucleotide reductase activity following induction of the nitrite-generating pathway in adenocarcinoma cells. J Biol Chem 265: 14143–14149PubMedGoogle Scholar
  45. Liew FY, Millott S, Parkinson C, Palmer RMJ, Moncada S (1990) Macrophage killing of Leishmania parasitein vivo is mediated by nitric oxide from L-arginine. J Immunol 144: 4794 4797Google Scholar
  46. Mackaness GB (1971) Resistance to intracellular infection. J Infect Dis 123: 439–445PubMedCrossRefGoogle Scholar
  47. Marietta MA, Yoon PS, Iyengar R, Leaf CD, Wishnock JS (1988) Macrophage oxidation of L—arginine to nitrite and nitrate: nitric oxide is an intermediate. Biochemistry 27: 8706–8711CrossRefGoogle Scholar
  48. Mayer J, Woods M, Vavrin Z, Hibbs JB Jr (1993) Gamma interferon-induced nitric oxide production reduces Chlamydia trachomatisinfectivity in McCoy cells. Infect Immun 61: 491–497PubMedGoogle Scholar
  49. Nathan CF, Hibbs JB Jr (1991) Role of nitric oxide synthesis in macrophage antimicrobial activity. Curr Opin Immunol 3: 65–70PubMedCrossRefGoogle Scholar
  50. Nussler A, Drapier J-C, Renia L, Pied S, Miltgen F, Gentilini M, Maxier D (1991) L-arginine-dependent destruction of intrahepatic malaria parasites in response to tumor necrosis factor and/or interleukin 6 stimulation. Eur J Immunol 21: 227–230PubMedCrossRefGoogle Scholar
  51. Palmer RMJ, Ashton DS, Moncada S (1988) Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 153: 1251–1256Google Scholar
  52. Pellat C, Henry Y, Drapier J-C (1990) IFN-7 activated macrophage: detection by electron paramagnetic resonance of complexes between L-arginine-derived nitric oxide and non-heme iron proteins. Biochem Biophys Res Commun 166: 119–125PubMedCrossRefGoogle Scholar
  53. Prütz W, Mönig H, Butler J, Land E (1985) Reactions of nitrogen dioxide in aqueous model systems: oxidation of tyrosine units in peptides and proteins. Arch Biochem Biophys 243: 125–134PubMedCrossRefGoogle Scholar
  54. Stamler JS, Singel DJ, Loscalzo (1992) Biochemistry of nitric oxide and its redoxactivated forms. Science 258: 1898–1902PubMedCrossRefGoogle Scholar
  55. Stuehr DJ, Nathan CF (1989) A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med 169: 1543–1555PubMedCrossRefGoogle Scholar
  56. Weinberg JB, Chapman HA Jr, Hibbs JB Jr (1978) Characterization of the effects of endotoxin on macrophage tumor cell killing. J Immunol 121: 72–80PubMedGoogle Scholar
  57. Wink DA, Kasprzak KS, Maragos CM, Elespuru RK, Misra J, Dunams TM, Cebula TA, Koch WH, Andrews AW, Allen JS, Keefer LK (1992) DNA deaminating ability and genotoxicity of nitric oxide and its progenitors. Science 254: 1001–1003CrossRefGoogle Scholar

Copyright information

© Springer-Verlag/Wien 1993

Authors and Affiliations

  • J. B. HibbsJr.
    • 1
  1. 1.VA Medical Center and Division of Infectious Diseases, School of MedicineThe University of UtahSalt Lake CityUSA

Personalised recommendations