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. 1996 Apr 15;97(8):1837–1843. doi: 10.1172/JCI118613

Nitric oxide inhibits viral replication in murine myocarditis.

C J Lowenstein 1, S L Hill 1, A Lafond-Walker 1, J Wu 1, G Allen 1, M Landavere 1, N R Rose 1, A Herskowitz 1
PMCID: PMC507251  PMID: 8621766

Abstract

Nitric oxide (NO) is a radical molecule that not only serves as a vasodilator and neurotransmitter but also acts as a cytotoxic effector molecule of the immune system. The inducible enzyme making NO, inducible NO synthase (iNOS), is transcriptionally activated by IFN-gamma and TNF-alpha, cytokines which are produced during viral infection. We show that iNOS is induced in mice infected with the Coxsackie B3 virus. Macrophages expressing iNOS are identified in the hearts and spleens of infected animals with an antibody raised against iNOS. Infected mice have increased titers of virus and a higher mortality when fed NOS inhibitors. Thus, viral infection induces iNOS in vivo, and NO inhibits viral replication. NO is a novel, nonspecific immune defense against viruses in vivo.

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Selected References

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  1. Adams L. B., Franzblau S. G., Vavrin Z., Hibbs J. B., Jr, Krahenbuhl J. L. L-arginine-dependent macrophage effector functions inhibit metabolic activity of Mycobacterium leprae. J Immunol. 1991 Sep 1;147(5):1642–1646. [PubMed] [Google Scholar]
  2. Balligand J. L., Ungureanu-Longrois D., Simmons W. W., Pimental D., Malinski T. A., Kapturczak M., Taha Z., Lowenstein C. J., Davidoff A. J., Kelly R. A. Cytokine-inducible nitric oxide synthase (iNOS) expression in cardiac myocytes. Characterization and regulation of iNOS expression and detection of iNOS activity in single cardiac myocytes in vitro. J Biol Chem. 1994 Nov 4;269(44):27580–27588. [PubMed] [Google Scholar]
  3. Beckerman K. P., Rogers H. W., Corbett J. A., Schreiber R. D., McDaniel M. L., Unanue E. R. Release of nitric oxide during the T cell-independent pathway of macrophage activation. Its role in resistance to Listeria monocytogenes. J Immunol. 1993 Feb 1;150(3):888–895. [PubMed] [Google Scholar]
  4. Bi Z., Reiss C. S. Inhibition of vesicular stomatitis virus infection by nitric oxide. J Virol. 1995 Apr;69(4):2208–2213. doi: 10.1128/jvi.69.4.2208-2213.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolaños J. P., Peuchen S., Heales S. J., Land J. M., Clark J. B. Nitric oxide-mediated inhibition of the mitochondrial respiratory chain in cultured astrocytes. J Neurochem. 1994 Sep;63(3):910–916. doi: 10.1046/j.1471-4159.1994.63030910.x. [DOI] [PubMed] [Google Scholar]
  6. Bredt D. S., Hwang P. M., Snyder S. H. Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature. 1990 Oct 25;347(6295):768–770. doi: 10.1038/347768a0. [DOI] [PubMed] [Google Scholar]
  7. Bredt D. S., Snyder S. H. Isolation of nitric oxide synthetase, a calmodulin-requiring enzyme. Proc Natl Acad Sci U S A. 1990 Jan;87(2):682–685. doi: 10.1073/pnas.87.2.682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brüne B., Lapetina E. G. Activation of a cytosolic ADP-ribosyltransferase by nitric oxide-generating agents. J Biol Chem. 1989 May 25;264(15):8455–8458. [PubMed] [Google Scholar]
  9. Brüne B., Lapetina E. G. Properties of a novel nitric oxide-stimulated ADP-ribosyltransferase. Arch Biochem Biophys. 1990 Jun;279(2):286–290. doi: 10.1016/0003-9861(90)90493-i. [DOI] [PubMed] [Google Scholar]
  10. Bukrinsky M. I., Nottet H. S., Schmidtmayerova H., Dubrovsky L., Flanagan C. R., Mullins M. E., Lipton S. A., Gendelman H. E. Regulation of nitric oxide synthase activity in human immunodeficiency virus type 1 (HIV-1)-infected monocytes: implications for HIV-associated neurological disease. J Exp Med. 1995 Feb 1;181(2):735–745. doi: 10.1084/jem.181.2.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Croen K. D. Evidence for antiviral effect of nitric oxide. Inhibition of herpes simplex virus type 1 replication. J Clin Invest. 1993 Jun;91(6):2446–2452. doi: 10.1172/JCI116479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Denis M. Interferon-gamma-treated murine macrophages inhibit growth of tubercle bacilli via the generation of reactive nitrogen intermediates. Cell Immunol. 1991 Jan;132(1):150–157. doi: 10.1016/0008-8749(91)90014-3. [DOI] [PubMed] [Google Scholar]
  13. Dighiero P., Reux I., Hauw J. J., Fillet A. M., Courtois Y., Goureau O. Expression of inducible nitric oxide synthase in cytomegalovirus-infected glial cells of retinas from AIDS patients. Neurosci Lett. 1994 Jan 17;166(1):31–34. doi: 10.1016/0304-3940(94)90833-8. [DOI] [PubMed] [Google Scholar]
  14. Egdell R. M., Siminiak T., Sheridan D. J. Modulation of neutrophil activity by nitric oxide during acute myocardial ischaemia and reperfusion. Basic Res Cardiol. 1994 Nov-Dec;89(6):499–509. doi: 10.1007/BF00794950. [DOI] [PubMed] [Google Scholar]
  15. Esumi H., Tannenbaum S. R. U.S.-Japan Cooperative Cancer Research Program: seminar on nitric oxide synthase and carcinogenesis. Cancer Res. 1994 Jan 1;54(1):297–301. [PubMed] [Google Scholar]
  16. 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]
  17. Godeny E. K., Gauntt C. J. Involvement of natural killer cells in coxsackievirus B3-induced murine myocarditis. J Immunol. 1986 Sep 1;137(5):1695–1702. [PubMed] [Google Scholar]
  18. Godeny E. K., Gauntt C. J. Murine natural killer cells limit coxsackievirus B3 replication. J Immunol. 1987 Aug 1;139(3):913–918. [PubMed] [Google Scholar]
  19. Granger D. L., Hibbs J. B., Jr, Broadnax L. M. Urinary nitrate excretion in relation to murine macrophage activation. Influence of dietary L-arginine and oral NG-monomethyl-L-arginine. J Immunol. 1991 Feb 15;146(4):1294–1302. [PubMed] [Google Scholar]
  20. Harris N., Buller R. M., Karupiah G. Gamma interferon-induced, nitric oxide-mediated inhibition of vaccinia virus replication. J Virol. 1995 Feb;69(2):910–915. doi: 10.1128/jvi.69.2.910-915.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Henke A., Mohr C., Sprenger H., Graebner C., Stelzner A., Nain M., Gemsa D. Coxsackievirus B3-induced production of tumor necrosis factor-alpha, IL-1 beta, and IL-6 in human monocytes. J Immunol. 1992 Apr 1;148(7):2270–2277. [PubMed] [Google Scholar]
  22. Henke A., Spengler H. P., Stelzner A., Nain M., Gemsa D. Lipopolysaccharide suppresses cytokine release from coxsackie virus-infected human monocytes. Res Immunol. 1992 Jan;143(1):65–70. doi: 10.1016/0923-2494(92)80081-u. [DOI] [PubMed] [Google Scholar]
  23. Herskowitz A., Wolfgram L. J., Rose N. R., Beisel K. W. Coxsackievirus B3 murine myocarditis: a pathologic spectrum of myocarditis in genetically defined inbred strains. J Am Coll Cardiol. 1987 Jun;9(6):1311–1319. doi: 10.1016/s0735-1097(87)80471-0. [DOI] [PubMed] [Google Scholar]
  24. Ignarro L. J. Biosynthesis and metabolism of endothelium-derived nitric oxide. Annu Rev Pharmacol Toxicol. 1990;30:535–560. doi: 10.1146/annurev.pa.30.040190.002535. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Kilbourn R. G., Klostergaard J., Lopez-Berestein G. Activated macrophages secrete a soluble factor that inhibits mitochondrial respiration of tumor cells. J Immunol. 1984 Nov;133(5):2577–2581. [PubMed] [Google Scholar]
  27. Kots AYa, Skurat A. V., Sergienko E. A., Bulargina T. V., Severin E. S. Nitroprusside stimulates the cysteine-specific mono(ADP-ribosylation) of glyceraldehyde-3-phosphate dehydrogenase from human erythrocytes. FEBS Lett. 1992 Mar 23;300(1):9–12. doi: 10.1016/0014-5793(92)80153-8. [DOI] [PubMed] [Google Scholar]
  28. Kwon N. S., Stuehr D. J., Nathan C. F. Inhibition of tumor cell ribonucleotide reductase by macrophage-derived nitric oxide. J Exp Med. 1991 Oct 1;174(4):761–767. doi: 10.1084/jem.174.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lane J. R., Neumann D. A., Lafond-Walker A., Herskowitz A., Rose N. R. Interleukin 1 or tumor necrosis factor can promote Coxsackie B3-induced myocarditis in resistant B10.A mice. J Exp Med. 1992 Apr 1;175(4):1123–1129. doi: 10.1084/jem.175.4.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lane J. R., Neumann D. A., Lafond-Walker A., Herskowitz A., Rose N. R. LPS promotes CB3-induced myocarditis in resistant B10.A mice. Cell Immunol. 1991 Aug;136(1):219–233. doi: 10.1016/0008-8749(91)90396-s. [DOI] [PubMed] [Google Scholar]
  31. Lane J. R., Neumann D. A., Lafond-Walker A., Herskowitz A., Rose N. R. Role of IL-1 and tumor necrosis factor in coxsackie virus-induced autoimmune myocarditis. J Immunol. 1993 Aug 1;151(3):1682–1690. [PubMed] [Google Scholar]
  32. Lefer A. M., Tsao P. S., Ma X. L., Sampath T. K. Anti-ischaemic and endothelial protective actions of recombinant human osteogenic protein (hOP-1). J Mol Cell Cardiol. 1992 Jun;24(6):585–593. doi: 10.1016/0022-2828(92)91043-5. [DOI] [PubMed] [Google Scholar]
  33. Lepoivre M., Fieschi F., Coves J., Thelander L., Fontecave M. Inactivation of ribonucleotide reductase by nitric oxide. Biochem Biophys Res Commun. 1991 Aug 30;179(1):442–448. doi: 10.1016/0006-291x(91)91390-x. [DOI] [PubMed] [Google Scholar]
  34. Liu R. H., Jacob J. R., Hotchkiss J. H., Cote P. J., Gerin J. L., Tennant B. C. Woodchuck hepatitis virus surface antigen induces nitric oxide synthesis in hepatocytes: possible role in hepatocarcinogenesis. Carcinogenesis. 1994 Dec;15(12):2875–2877. doi: 10.1093/carcin/15.12.2875. [DOI] [PubMed] [Google Scholar]
  35. Lowenstein C. J., Alley E. W., Raval P., Snowman A. M., Snyder S. H., Russell S. W., Murphy W. J. Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9730–9734. doi: 10.1073/pnas.90.20.9730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lowenstein C. J., Glatt C. S., Bredt D. S., Snyder S. H. Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6711–6715. doi: 10.1073/pnas.89.15.6711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lowenstein C. J., Snyder S. H. Nitric oxide, a novel biologic messenger. Cell. 1992 Sep 4;70(5):705–707. doi: 10.1016/0092-8674(92)90301-r. [DOI] [PubMed] [Google Scholar]
  38. Lyons C. R., Orloff G. J., Cunningham J. M. Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophage cell line. J Biol Chem. 1992 Mar 25;267(9):6370–6374. [PubMed] [Google Scholar]
  39. Ma X. L., Weyrich A. S., Lefer D. J., Lefer A. M. Diminished basal nitric oxide release after myocardial ischemia and reperfusion promotes neutrophil adherence to coronary endothelium. Circ Res. 1993 Feb;72(2):403–412. doi: 10.1161/01.res.72.2.403. [DOI] [PubMed] [Google Scholar]
  40. MacMicking J. D., Nathan C., Hom G., Chartrain N., Fletcher D. S., Trumbauer M., Stevens K., Xie Q. W., Sokol K., Hutchinson N. Altered responses to bacterial infection and endotoxic shock in mice lacking inducible nitric oxide synthase. Cell. 1995 May 19;81(4):641–650. doi: 10.1016/0092-8674(95)90085-3. [DOI] [PubMed] [Google Scholar]
  41. Mannick J. B., Asano K., Izumi K., Kieff E., Stamler J. S. Nitric oxide produced by human B lymphocytes inhibits apoptosis and Epstein-Barr virus reactivation. Cell. 1994 Dec 30;79(7):1137–1146. doi: 10.1016/0092-8674(94)90005-1. [DOI] [PubMed] [Google Scholar]
  42. Marletta M. A. Nitric oxide synthase: aspects concerning structure and catalysis. Cell. 1994 Sep 23;78(6):927–930. doi: 10.1016/0092-8674(94)90268-2. [DOI] [PubMed] [Google Scholar]
  43. Mateo R. B., Reichner J. S., Mastrofrancesco B., Kraft-Stolar D., Albina J. E. Impact of nitric oxide on macrophage glucose metabolism and glyceraldehyde-3-phosphate dehydrogenase activity. Am J Physiol. 1995 Mar;268(3 Pt 1):C669–C675. doi: 10.1152/ajpcell.1995.268.3.C669. [DOI] [PubMed] [Google Scholar]
  44. Moncada S., Palmer R. M., Higgs E. A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991 Jun;43(2):109–142. [PubMed] [Google Scholar]
  45. Nathan C., Xie Q. W. Nitric oxide synthases: roles, tolls, and controls. Cell. 1994 Sep 23;78(6):915–918. doi: 10.1016/0092-8674(94)90266-6. [DOI] [PubMed] [Google Scholar]
  46. Nguyen T., Brunson D., Crespi C. L., Penman B. W., Wishnok J. S., Tannenbaum S. R. DNA damage and mutation in human cells exposed to nitric oxide in vitro. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3030–3034. doi: 10.1073/pnas.89.7.3030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Niu X. F., Smith C. W., Kubes P. Intracellular oxidative stress induced by nitric oxide synthesis inhibition increases endothelial cell adhesion to neutrophils. Circ Res. 1994 Jun;74(6):1133–1140. doi: 10.1161/01.res.74.6.1133. [DOI] [PubMed] [Google Scholar]
  48. Pinsky D. J., Cai B., Yang X., Rodriguez C., Sciacca R. R., Cannon P. J. The lethal effects of cytokine-induced nitric oxide on cardiac myocytes are blocked by nitric oxide synthase antagonism or transforming growth factor beta. J Clin Invest. 1995 Feb;95(2):677–685. doi: 10.1172/JCI117713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Rice W. G., Schaeffer C. A., Harten B., Villinger F., South T. L., Summers M. F., Henderson L. E., Bess J. W., Jr, Arthur L. O., McDougal J. S. Inhibition of HIV-1 infectivity by zinc-ejecting aromatic C-nitroso compounds. Nature. 1993 Feb 4;361(6411):473–475. doi: 10.1038/361473a0. [DOI] [PubMed] [Google Scholar]
  50. Russell M. E., Wallace A. F., Wyner L. R., Newell J. B., Karnovsky M. J. Upregulation and modulation of inducible nitric oxide synthase in rat cardiac allografts with chronic rejection and transplant arteriosclerosis. Circulation. 1995 Aug 1;92(3):457–464. doi: 10.1161/01.cir.92.3.457. [DOI] [PubMed] [Google Scholar]
  51. Sentman C. L., Hackett J., Jr, Kumar V., Bennett M. Identification of a subset of murine natural killer cells that mediates rejection of Hh-1d but not Hh-1b bone marrow grafts. J Exp Med. 1989 Jul 1;170(1):191–202. doi: 10.1084/jem.170.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Springer T. A. Monoclonal antibody analysis of complex biological systems. Combination of cell hybridization and immunoadsorbents in a novel cascade procedure and its application to the macrophage cell surface. J Biol Chem. 1981 Apr 25;256(8):3833–3839. [PubMed] [Google Scholar]
  53. Springer T., Galfrè G., Secher D. S., Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978 Aug;8(8):539–551. doi: 10.1002/eji.1830080802. [DOI] [PubMed] [Google Scholar]
  54. Stamler J. S. Redox signaling: nitrosylation and related target interactions of nitric oxide. Cell. 1994 Sep 23;78(6):931–936. doi: 10.1016/0092-8674(94)90269-0. [DOI] [PubMed] [Google Scholar]
  55. Ungureanu-Longrois D., Balligand J. L., Simmons W. W., Okada I., Kobzik L., Lowenstein C. J., Kunkel S. L., Michel T., Kelly R. A., Smith T. W. Induction of nitric oxide synthase activity by cytokines in ventricular myocytes is necessary but not sufficient to decrease contractile responsiveness to beta-adrenergic agonists. Circ Res. 1995 Sep;77(3):494–502. doi: 10.1161/01.res.77.3.494. [DOI] [PubMed] [Google Scholar]
  56. Wei X. Q., Charles I. G., Smith A., Ure J., Feng G. J., Huang F. P., Xu D., Muller W., Moncada S., Liew F. Y. Altered immune responses in mice lacking inducible nitric oxide synthase. Nature. 1995 Jun 1;375(6530):408–411. doi: 10.1038/375408a0. [DOI] [PubMed] [Google Scholar]
  57. Welsh N., Eizirik D. L., Bendtzen K., Sandler S. Interleukin-1 beta-induced nitric oxide production in isolated rat pancreatic islets requires gene transcription and may lead to inhibition of the Krebs cycle enzyme aconitase. Endocrinology. 1991 Dec;129(6):3167–3173. doi: 10.1210/endo-129-6-3167. [DOI] [PubMed] [Google Scholar]
  58. Wink D. A., Kasprzak K. S., Maragos C. M., Elespuru R. K., Misra M., Dunams T. M., Cebula T. A., Koch W. H., Andrews A. W., Allen J. S. DNA deaminating ability and genotoxicity of nitric oxide and its progenitors. Science. 1991 Nov 15;254(5034):1001–1003. doi: 10.1126/science.1948068. [DOI] [PubMed] [Google Scholar]
  59. Wolfgram L. J., Beisel K. W., Herskowitz A., Rose N. R. Variations in the susceptibility to Coxsackievirus B3-induced myocarditis among different strains of mice. J Immunol. 1986 Mar 1;136(5):1846–1852. [PubMed] [Google Scholar]
  60. Worrall N. K., Lazenby W. D., Misko T. P., Lin T. S., Rodi C. P., Manning P. T., Tilton R. G., Williamson J. R., Ferguson T. B., Jr Modulation of in vivo alloreactivity by inhibition of inducible nitric oxide synthase. J Exp Med. 1995 Jan 1;181(1):63–70. doi: 10.1084/jem.181.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Xie Q. W., Cho H. J., Calaycay J., Mumford R. A., Swiderek K. M., Lee T. D., Ding A., Troso T., Nathan C. Cloning and characterization of inducible nitric oxide synthase from mouse macrophages. Science. 1992 Apr 10;256(5054):225–228. doi: 10.1126/science.1373522. [DOI] [PubMed] [Google Scholar]
  62. Xie Q. W., Whisnant R., Nathan C. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide. J Exp Med. 1993 Jun 1;177(6):1779–1784. doi: 10.1084/jem.177.6.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Yang X., Chowdhury N., Cai B., Brett J., Marboe C., Sciacca R. R., Michler R. E., Cannon P. J. Induction of myocardial nitric oxide synthase by cardiac allograft rejection. J Clin Invest. 1994 Aug;94(2):714–721. doi: 10.1172/JCI117390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Zhang J., Snyder S. H. Purification of a nitric oxide-stimulated ADP-ribosylated protein using biotinylated beta-nicotinamide adenine dinucleotide. Biochemistry. 1993 Mar 9;32(9):2228–2233. doi: 10.1021/bi00060a014. [DOI] [PubMed] [Google Scholar]

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