Nitric Oxide in TMEV

  • Emilia L. Oleszak
  • Jacek Kuzmak
  • Arun Varadhachary
  • Christos D. Katsetos


We and others have previously investigated the role of inducible nitric oxide synthase (iNOS) on early acute and late chronic demyelinating disease induced by Theiler’s Murine Encephalomyelitis Virus (TMEV). Infection of susceptible SJL mice with this virus serves as an excellent model of virus-induced demyelinating disease, such as multiple sclerosis (MS). iNOS transcripts and protein were detected in brains and spinal cords of TMEV-infected SJL mice during early acute disease, which resembles polioencephalomyelitis. Similar level of expression of iNOS has been found in resistant B6 mice, which develop only early acute disease. Weak iNOS staining was detected in reactive astrocytes and in leptomeningeal infiltrates in TMEV-infected SJL mice at 42 days post infection (p.i.), corresponding to early phase of chronic demyelinating disease, but not at 66 and 180 days p.i. corresponding to advanced and terminal stages of the disease, respectively. Results from other laboratories demonstrated that, blocking of NO by treatment of TMEV-infected SJL mice with amino guanidine (AG), a specific inhibitor of NO resulted in delay of late chronic demyelinating disease. However this protective effect of NO inhibitor depended on the temporal phase of the disease, type of cells expressing iNOS and the time of administration of AG. The results from our laboratory suggests that NO expressed during early acute disease is beneficial to the host through induction of apoptosis of infiltrating T cells and resolution of encephalitis, but its role in myelin/oligodendrocytes damage during late chronic demyelinating disease is not clear and it may depend on availability of superoxide and formation of peroxynitrite.

Key words

nitric oxide TMEV multiple sclerosis 


  1. 1.
    Oleszak EL, Kuzmak J, Good RA, Platsoucas. CD. Immunobiology of TMEV infection. Immunology of Theiler’s murine encephalomyelitis virus infection. Immunologic Res. 1995;1(13–33.).Google Scholar
  2. 2.
    Dal Canto M., Lipton. HL. Multiple sclerosis. Animal model: Theiler’s virus infection in mice. Am. J. Pathol. 1977;4(497–500.).Google Scholar
  3. 3.
    Levy M., Aubert C., Brahic. M. Theiler(s virus replication in brain macrophages cultured in vitro. J. Virol. 1992;3(3188–3193.).Google Scholar
  4. 4.
    Lipton HL. Characterization of the TO strains of Theiler’s mouse encephalomyelitis viuses. Infect. Immun. 1978;1(869–872.).Google Scholar
  5. 5.
    Lipton HL. Theiler’s virus infection in mice: an unusual biphasic disease process leading to demyelination. Infect. Immunol. 1975;0(1147–1155.).Google Scholar
  6. 6.
    Lindsley MD, Rodriguez. M. Characterization of the inflammatory response in the central nervous system of mice susceptible or resistant to demyelination by Theiler(s virus. J. Immunol. 1989;6(2677–2682.).Google Scholar
  7. 7.
    Tsunoda I, Fujinami RS. Two models for multiple sclerosis: experimental allergic encephalomyelitis and Theiler’s murine encephalomyelitis virus. J Neuropathol Exp Neurol. 1996;55(6):673–86.PubMedCrossRefGoogle Scholar
  8. 8.
    Brahic M, Stroop WG, Baringer. JR. Theiler’s virus persists in glial cells during demyelinating disease. Cell. 1981;1(123–128.).CrossRefGoogle Scholar
  9. 9.
    Rodriguez M, Oleszak E, Leibowitz J. Theiler’s murine encepha-lomyelitis: a model of demyelination and persistence of virus. CRC Crit. Rev. Immunol. 1987;7(325–365.).Google Scholar
  10. 10.
    Theiler M. Spontaneous encephalomyelitis of mice: a new virus disease. J. Exp. Med. 1937;3(705–719.).CrossRefGoogle Scholar
  11. 11.
    Kim BS, Palna JP. Immune mechanisms of Theiler’s virus-induced demyelination. Exp Mol Med. 1999;31(3):115–21.PubMedGoogle Scholar
  12. 12.
    Martin R, McFarland H.Immunology of Multiple Sclerosis and experimental allergic encephalomyelitis. 1st ed London: Chapman & Hall Medical; 1997. (Raine CS, McFarland HF, Tourtellotte WW, eds. Multiple sclerosis: clinical and pathogenic basis).Google Scholar
  13. 13.
    Brosnan CF, Raine CS. Mechanisms of immune injury in multiple sclerosis. Brain Pathol. 1996;6(3):243–57.PubMedGoogle Scholar
  14. 14.
    Wekerle H.Immunology of Multiple Sclerosis. 3rd ed London: Churchill Livingstone; 1998. (Compston A, ed. McAlpine’s multiple sclerosis).Google Scholar
  15. 15.
    Prineas J, McDonald W.Demyelinating Disease. 6th ed London, New York: Oxford University Press; 1997. (Greenfield JG, Lantos PL, Graham DI, eds. Greenfield’s Neuropathology).Google Scholar
  16. 16.
    Selmaj K, Raine CS, Farooq M, Norton WT, Brosnan CF. Cytokine cytotoxicity against oligodendrocytes. Apoptosis induced by lymphotoxin. J Immunol. 1991;147(5):1522–9.PubMedGoogle Scholar
  17. 17.
    Marlena MA. Nitric oxide synthase structure and mechanism. J. Biol. Chem. 1993;11(12231–12234.).Google Scholar
  18. 18.
    Nathan C., Xie. QW. Regulation of biosynthesis of nitric oxide. J. Biol. Chem. 1994;11(13725–13728.).Google Scholar
  19. 19.
    Bredt DS. Endogenous nitric oxide synthesis: biological functions and pathophysiology. Free Radic Res. 1999;31(6):577–96.PubMedCrossRefGoogle Scholar
  20. 20.
    Knowles RG, Moncada S. Nitric oxide synthases in mammals. Biochem J. 1994;298 (Pt 2):249–58.PubMedGoogle Scholar
  21. 21.
    Marietta MA, Yocr PS, Lyengar R, Leaf CD, Wlshnok. JS. Macrophage oxidation of L-arginine to nitrite and nitrate: nitric oxide is an intermediate. Biochemistry. 1988;1(8706–8711.).CrossRefGoogle Scholar
  22. 22.
    Nussler AK, Billiar. TR. Inflammation, immunoregulation, and inducible nitric oxide synthase. J. Leukocyte Biol. 1993;2(171–178.).Google Scholar
  23. 23.
    Bogdan C., Rollinghoff M., Diefenbach A. The role of nitric oxide in innate immunity. Immunol Rev. 2000;173:17–26.PubMedCrossRefGoogle Scholar
  24. 24.
    Di Rosa F, Barnaba V. Persisting viruses and chronic inflammation: understanding their relation to autoimmunity. Immunol Rev. 1998;164:17–27.PubMedCrossRefGoogle Scholar
  25. 25.
    Gazzinelli RT, Oswald IP, Hieny S, James SL, Sher A. The microbicidal activity of interferon-gamma-treated macrophages against Trypanosoma cruzi involves an L-arginine-dependent, nitrogen oxide-mediated mechanism inhibitable by interleukin-10 and transforming growth factor-beta. Eur J Immunol. 1992;22(10):2501–6.PubMedCrossRefGoogle Scholar
  26. 26.
    Chan J, Tanaka K, Carroll D, Flynn J, Bloom BR. Effects of nitric oxide synthase inhibitors on murine infection with Mycobacterium tuberculosis. Infect Immun. 1995;63(2):736–40.PubMedGoogle Scholar
  27. 27.
    Xie Q, Kawakami K, Kudeken N, Zhang T, Qureshi MH, Saito A. Different susceptibility of three clinically isolated strains of Cryptococcus neoformans to the fungicidal effects of reactive nitrogen and oxygen intermediates: possible relationships with virulence. Microbiol Immunol. 1997;41(9):725–31.PubMedGoogle Scholar
  28. 28.
    Shi FD, Flodstrom M, Kim SH, et al. Control of the autoimmune response by type 2 nitric oxide synthase. J Immunol. 2001;167(5):3000–6.PubMedGoogle Scholar
  29. 29.
    Kolb H, Kolb-Bachofen. V. Nitric oxide; A pathogenic factor in autoimmunity. Immunol. Today. 1992;1(157–160.).CrossRefGoogle Scholar
  30. 30.
    Kolb H, Kolb-Bachofen V. Nitric oxide in autoimmune disease: cytotoxic or regulatory mediator? Immunol Today. 1998;19(12):556–61.PubMedCrossRefGoogle Scholar
  31. 31.
    Moulian N, Truffault F, Gaudry-Talarmain YM, Serraf A, Berrih-Aknin S. In vivo and in vitro apoptosis of human thymocytes are associated with nitrotyrosine formation. Blood. 2001;97(11):3521–30.PubMedCrossRefGoogle Scholar
  32. 32.
    Marks-Konczalik J, Chu SC, Moss J. Cytokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor kappaB-binding sites. J Biol Chem. 1998;273(35):22201–8.PubMedCrossRefGoogle Scholar
  33. 33.
    Nathan C., Xie QW. Nitric oxide synthases: roles, tolls, and controls. Cell. 1994;78(6):915–8.PubMedCrossRefGoogle Scholar
  34. 34.
    Dawson VL, Dawson TM, London ED, Bredt DS, Snyder. SH. Nitric oxide mediates glutamate neurotoxicity in primary cultures. Proc. Natl. Acad. Sci. USA. 1991;4(6368–6371.).CrossRefGoogle Scholar
  35. 35.
    Merrill JE, Ignarro LJ, Sherman MP, Melinek J, Lane. TE. Microglial cell cytotoxicity of oligodentrocytes is mediated through nitric oxide. J. Immunol. 1993;6(2132–2141.).Google Scholar
  36. 36.
    Mitrovic B, Ignarro LJ, Montestruque S, Smoll A, Merrill. JE. Nitric oxide as a potential pathological mechanism in demyelination: its differential effects on primary glial cells in vitro. Neuroscience. 1994;3(575–585.).CrossRefGoogle Scholar
  37. 37.
    Wink DA, Kasprzak KS, Maragos CM, et al. DNA deaminating ability and genotoxicity of nitric oxide and its progenitors. Science. 1991;11(1001–1003.).CrossRefGoogle Scholar
  38. 38.
    Stuehr DJ, Nathan CF. Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med. 1989; 169(5): 1543–55.PubMedCrossRefGoogle Scholar
  39. 39.
    Beckman JS, Crow. JP. Pathological implications of nitric oxide, superoxide and peroxynitrite formation. Biochem. Soc. Trans. 1993;1(330–334.).Google Scholar
  40. 40.
    Lipton SA, Choi YB, Pan ZH, et al. A redox-based mechanism for the neuroprotective and neurodestructive effects of nitric oxide and related nitrosocompounds. Nature. 1993;15(626–631.).CrossRefGoogle Scholar
  41. 41.
    Zielasek J, Tausch M, Toyka KV, Hartung. HP. Production of nitrite by neonatal rat microglial cells/brain macrophages. Cell. Immunol. 1992;6(111–120.).CrossRefGoogle Scholar
  42. 42.
    Hunter MI, Nlemadim BC, Davidson DL. Lipid peroxidation products and antioxidant proteins in plasma and cerebrospinal fluid from multiple sclerosis patients. Neurochem Res. 1985;10(12): 1645–52.PubMedCrossRefGoogle Scholar
  43. 43.
    Liu JS, Zhao ML, Brosnan CF, Lee SC. Expression of inducible nitric oxide synthase and nitrotyrosine in multiple sclerosis lesions. Am J Pathol. 2001; 158(6):2057–66.PubMedGoogle Scholar
  44. 44.
    Liu B, Gao HM, Wang JY, Jeohn GH, Cooper CL, Hong JS. Role of nitric oxide in inflammation-mediated neurodegeneration. Ann N Y Acad Sci. 2002;962:318–31.PubMedGoogle Scholar
  45. 45.
    Oleszak EL, Zaczynska E, Bhattacharjee M, Butunoi C., Legido A, Katsetos CD. Inducible nitric oxide synthase and nitrotyrosine are found in monocytes/macrophages and/or astrocytes in acute, but not in chronic, multiple sclerosis. Clin Diagn Lab Immunol. 1998;5(4):438–45.PubMedGoogle Scholar
  46. 46.
    Bagasra O, Michaels FH, Zheng YM, et al. Activation of the inducible form of nitric oxide synthase in the brains of patients with multiple sclerosis. Proc. Natl. Acad. Sci. USA. 1995;4(12041–12045.).CrossRefGoogle Scholar
  47. 47.
    Bö L, Dawson TM, Wesselingh S, et al. Induction of nitric oxide synthase in demyelinating regions of multiple sclerosis brains. Ann. Neurol. 1994;2(778–786.).CrossRefGoogle Scholar
  48. 48.
    Galea E, Reis DJ, Feinstein. DL. Cloning and expression of inducible nitric oxide synthase from rat astrocytes. J. Neurosci. Res. 1994;2(406–414).CrossRefGoogle Scholar
  49. 49.
    Hewett SJ, Corbett JA, McDaniel ML, Choi. DW. IFN-? and IL-1β induce nitric oxide formation from primary mouse astrocytes. Neurosci. Lett. 1993;7(229–232.).CrossRefGoogle Scholar
  50. 50.
    Minc-Golomb D, Tsarfaty I, Schwartz. JP. Expression of inducible nitric oxide synthase by neurones following exposure to endotoxin and cytokine. Br. J. Pharmacol. 1994;5(720–722.).Google Scholar
  51. 51.
    Van Dam A-M, Bauer J, Man-A-Hing WKH, Marquette C., Tilders FJH, Berkenbosch. F. Appearance of inducible nitric oxide synthase in the rat central nervous system after rabies virus infection and during experimental allergic encephalomyelitis but not after peripheral administration of endotoxin. J. Neurosci. Res. 1995;2(251–260.).Google Scholar
  52. 52.
    Reiss CS, Komatsu T. Does nitric oxide play a critical role in viral infections? J Virol. 1998;72(6):4547–51.PubMedGoogle Scholar
  53. 53.
    Wu GF, Pewe L, Perlman S. Coronavirus-induced demyelination occurs in the absence of inducible nitric oxide synthase. J Virol. 2000;74(16):7683–6.PubMedCrossRefGoogle Scholar
  54. 54.
    Lane TE, Paoletti AD, Buchmeier MJ. Disassociation between the in vitro and in vivo effects of nitric oxide on aneurotropic murine coronavirus. J Virol. 1997;71(3):2202–10.PubMedGoogle Scholar
  55. 55.
    Bartholdy C, Nansen A, Christensen JE, Marker O, Thomsen AR. Inducible nitric-oxide synthase plays a minimal role in lymphocytic choriomeningitis virus-induced, T cell-mediated protective immunity and immunopathology. J Gen Virol. 1999;80(Pt 11):2997–3005.PubMedGoogle Scholar
  56. 56.
    Schoneboom BA, Catlin KM, Marty AM, Grieder FB. Inflammation is a component of neurodegeneration in response to Venezuelan equine encephalitis virus infection in mice. J Neuroimmunol. 2000;109(2): 132–46.PubMedCrossRefGoogle Scholar
  57. 57.
    Saxena SK, Mathur A, Srivastava RC. Induction of nitric oxide synthase during Japanese encephalitis virus infection: evidence of protective role. Arch Biochem Biophys. 2001;391(1):1–7.PubMedCrossRefGoogle Scholar
  58. 58.
    Kreil TR, Eibl MM. Nitric oxide and viral infection: NO antiviral activity against a flavivirus in vitro, and evidence for contribution to pathogenesis in experimental infection in vivo. Virology. 1996;219(1):304–6.PubMedCrossRefGoogle Scholar
  59. 59.
    Koprowski H, Zhen YM, Heber-Katz E, et al. In vivo expression of inducible nitric oxide synthase in experimentally induced neurologic diseases. Proc. Natl. Acad. Sci. 1993;4(3024–3027.).CrossRefGoogle Scholar
  60. 60.
    Torre D, Pugliese A, Speranza F. Role of nitric oxide in HIV-1 infection: friend or foe? Lancet Infect Dis. 2002;2(5):273–80.PubMedCrossRefGoogle Scholar
  61. 61.
    O1eszak EL, Katsetos CD, Kuzmak J, Varadhachary A. Inducible nitric oxide synthase in Theiler’s murine encephalomyelitis virus infection. J Virol. 1997;71(4):3228–35.Google Scholar
  62. 62.
    Xiao BG, Xu LY, Yang JS, Huang YM, Link H. An alternative pathway of nitric oxide production by rat astrocytes requires specific antigen and T cell contact. Neurosci Lett. 2000;283(1):53–6.PubMedCrossRefGoogle Scholar
  63. 63.
    Molina-Holgado E, Arevalo-Martin A, Castrillo A, Bosca L, Vela JM, Guaza C. Interleukin-4 and interleukin-10 modulate nuclear factor kappaB activity and nitric oxide synthase-2 expression in Theiler’s virus-infected brain astrocytes. J Neurochem. 2002;81(6):1242–52.PubMedCrossRefGoogle Scholar
  64. 64.
    Chang JR, Zaczynska E, Katsetos CD, Platsoucas CD, Oleszak EL. Differential expression of TGF-beta, IL-2, and other cytokines in the CNS of Theiler’s murine encephalomyelitis virus-infected susceptible and resistant strains of mice. Virology. 2000;278(2):346–60.PubMedCrossRefGoogle Scholar
  65. 65.
    WilIenborg DO, Staykova MA, Cowden WB. Our shifting understanding of the role of nitric oxide in autoimmune encephalomyelitis: a review. J Neuroimmunol. 1999;100(1–2):21–35.CrossRefGoogle Scholar
  66. 66.
    Rose JW, Hill KE, Wada Y, et al. Nitric oxide synthase inhibitor, aminoguanidine, reduces inflammation and demyelination produced by Theiler’s virus infection. J Neuroimmunol. 1998;81(1–2):82–9.PubMedCrossRefGoogle Scholar
  67. 67.
    Iwahashi T, Inoue A, Koh CS, Shin TK, Kim BS. Expression and potential role of inducible nitric oxide synthase in the central nervous system of Theiler’s murine encephalomyelitis virus-induced demyelinating disease. Cell Immunol. 1999;194(2): 186–93.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Emilia L. Oleszak
    • 1
  • Jacek Kuzmak
    • 1
  • Arun Varadhachary
    • 2
  • Christos D. Katsetos
    • 3
  1. 1.Fels Institute for Cancer Research and Molecular Biology and Department of Anatomy and Cell BiologyUSA
  2. 2.Department of Microbiology and ImmunologyTemple University School of MedicineUSA
  3. 3.College of Medicine and St. Christopher’s Hospital for ChildrenDrexel UniversityPhiladelphia

Personalised recommendations