Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1996 Mar;70(3):1828–1835. doi: 10.1128/jvi.70.3.1828-1835.1996

Alphavirus-induced apoptosis in mouse brains correlates with neurovirulence.

J Lewis 1, S L Wesselingh 1, D E Griffin 1, J M Hardwick 1
PMCID: PMC190010  PMID: 8627707

Abstract

Sindbis virus induces apoptotic cell death in cultured cell lines, raising the possibility that apoptosis of infected neurons and other target cells in vivo may contribute to the resulting disease and mortality. To investigate the role of apoptosis in Sindbis virus pathogenesis, infected mouse brains were assayed by the in situ terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling technique and for DNA ladder formation. Infection with recombinant Sindbis virus strain 633 resulted in widespread apoptosis in newborn mouse brains and spinal cords, but few apoptotic cells were observed following infection of 2-week-old animals. This finding correlates with the age-dependent mortality observed in mice. The more neurovirulent virus TE, which differs from 633 by a single amino acid in the E2 glycoprotein, induced significant apoptosis in brains and spinal cords of 2-week-old animals, consistent with its ability to cause fatal disease in older animals. Double-labeling experiments demonstrated that the apoptotic cells were also infected with Sindbis virus. Thus, Sindbis virus-induced apoptosis appears to be a result of virus infection and is likely to reflect pathogenic mechanisms for other viruses.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ameisen J. C., Estaquier J., Idziorek T., De Bels F. The relevance of apoptosis to AIDS pathogenesis. Trends Cell Biol. 1995 Jan;5(1):27–32. doi: 10.1016/s0962-8924(00)88933-3. [DOI] [PubMed] [Google Scholar]
  2. Clem R. J., Miller L. K. Apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. J Virol. 1993 Jul;67(7):3730–3738. doi: 10.1128/jvi.67.7.3730-3738.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clem R. J., Miller L. K. Control of programmed cell death by the baculovirus genes p35 and iap. Mol Cell Biol. 1994 Aug;14(8):5212–5222. doi: 10.1128/mcb.14.8.5212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cowan W. M., Fawcett J. W., O'Leary D. D., Stanfield B. B. Regressive events in neurogenesis. Science. 1984 Sep 21;225(4668):1258–1265. doi: 10.1126/science.6474175. [DOI] [PubMed] [Google Scholar]
  5. Ellis R. E., Jacobson D. M., Horvitz H. R. Genes required for the engulfment of cell corpses during programmed cell death in Caenorhabditis elegans. Genetics. 1991 Sep;129(1):79–94. doi: 10.1093/genetics/129.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Esolen L. M., Park S. W., Hardwick J. M., Griffin D. E. Apoptosis as a cause of death in measles virus-infected cells. J Virol. 1995 Jun;69(6):3955–3958. doi: 10.1128/jvi.69.6.3955-3958.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Estaquier J., Idziorek T., de Bels F., Barré-Sinoussi F., Hurtrel B., Aubertin A. M., Venet A., Mehtali M., Muchmore E., Michel P. Programmed cell death and AIDS: significance of T-cell apoptosis in pathogenic and nonpathogenic primate lentiviral infections. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9431–9435. doi: 10.1073/pnas.91.20.9431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Estus S., Zaks W. J., Freeman R. S., Gruda M., Bravo R., Johnson E. M., Jr Altered gene expression in neurons during programmed cell death: identification of c-jun as necessary for neuronal apoptosis. J Cell Biol. 1994 Dec;127(6 Pt 1):1717–1727. doi: 10.1083/jcb.127.6.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferrer I., Tortosa A., Condom E., Blanco R., Macaya A., Planas A. Increased expression of bcl-2 immunoreactivity in the developing cerebral cortex of the rat. Neurosci Lett. 1994 Sep 26;179(1-2):13–16. doi: 10.1016/0304-3940(94)90923-7. [DOI] [PubMed] [Google Scholar]
  10. Fesq H., Bacher M., Nain M., Gemsa D. Programmed cell death (apoptosis) in human monocytes infected by influenza A virus. Immunobiology. 1994 Feb;190(1-2):175–182. doi: 10.1016/S0171-2985(11)80292-5. [DOI] [PubMed] [Google Scholar]
  11. Fleming J. O. Viral neurovirulence. Lab Invest. 1988 May;58(5):481–483. [PubMed] [Google Scholar]
  12. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gelbard H. A., James H. J., Sharer L. R., Perry S. W., Saito Y., Kazee A. M., Blumberg B. M., Epstein L. G. Apoptotic neurons in brains from paediatric patients with HIV-1 encephalitis and progressive encephalopathy. Neuropathol Appl Neurobiol. 1995 Jun;21(3):208–217. doi: 10.1111/j.1365-2990.1995.tb01052.x. [DOI] [PubMed] [Google Scholar]
  14. Gold R., Schmied M., Giegerich G., Breitschopf H., Hartung H. P., Toyka K. V., Lassmann H. Differentiation between cellular apoptosis and necrosis by the combined use of in situ tailing and nick translation techniques. Lab Invest. 1994 Aug;71(2):219–225. [PubMed] [Google Scholar]
  15. Griffin D. E., Levine B., Tyor W. R., Tucker P. C., Hardwick J. M. Age-dependent susceptibility to fatal encephalitis: alphavirus infection of neurons. Arch Virol Suppl. 1994;9:31–39. doi: 10.1007/978-3-7091-9326-6_4. [DOI] [PubMed] [Google Scholar]
  16. Hershberger P. A., Dickson J. A., Friesen P. D. Site-specific mutagenesis of the 35-kilodalton protein gene encoded by Autographa californica nuclear polyhedrosis virus: cell line-specific effects on virus replication. J Virol. 1992 Sep;66(9):5525–5533. doi: 10.1128/jvi.66.9.5525-5533.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hinshaw V. S., Olsen C. W., Dybdahl-Sissoko N., Evans D. Apoptosis: a mechanism of cell killing by influenza A and B viruses. J Virol. 1994 Jun;68(6):3667–3673. doi: 10.1128/jvi.68.6.3667-3673.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hockenbery D. M., Zutter M., Hickey W., Nahm M., Korsmeyer S. J. BCL2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6961–6965. doi: 10.1073/pnas.88.16.6961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ishii H. H., Gobé G. C. Epstein-Barr virus infection is associated with increased apoptosis in untreated and phorbol ester-treated human Burkitt's lymphoma (AW-Ramos) cells. Biochem Biophys Res Commun. 1993 May 14;192(3):1415–1423. doi: 10.1006/bbrc.1993.1574. [DOI] [PubMed] [Google Scholar]
  20. Jackson A. C., Moench T. R., Trapp B. D., Griffin D. E. Basis of neurovirulence in Sindbis virus encephalomyelitis of mice. Lab Invest. 1988 May;58(5):503–509. [PubMed] [Google Scholar]
  21. Jeurissen S. H., Wagenaar F., Pol J. M., van der Eb A. J., Noteborn M. H. Chicken anemia virus causes apoptosis of thymocytes after in vivo infection and of cell lines after in vitro infection. J Virol. 1992 Dec;66(12):7383–7388. doi: 10.1128/jvi.66.12.7383-7388.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnson P. A., Miyanohara A., Levine F., Cahill T., Friedmann T. Cytotoxicity of a replication-defective mutant of herpes simplex virus type 1. J Virol. 1992 May;66(5):2952–2965. doi: 10.1128/jvi.66.5.2952-2965.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kawanishi M. Epstein-Barr virus induces fragmentation of chromosomal DNA during lytic infection. J Virol. 1993 Dec;67(12):7654–7658. doi: 10.1128/jvi.67.12.7654-7658.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koga Y., Tanaka K., Lu Y. Y., Oh-Tsu M., Sasaki M., Kimura G., Nomoto K. Priming of immature thymocytes to CD3-mediated apoptosis by infection with murine cytomegalovirus. J Virol. 1994 Jul;68(7):4322–4328. doi: 10.1128/jvi.68.7.4322-4328.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Krajewski S., Krajewska M., Shabaik A., Miyashita T., Wang H. G., Reed J. C. Immunohistochemical determination of in vivo distribution of Bax, a dominant inhibitor of Bcl-2. Am J Pathol. 1994 Dec;145(6):1323–1336. [PMC free article] [PubMed] [Google Scholar]
  26. Laurent-Crawford A. G., Krust B., Muller S., Rivière Y., Rey-Cuillé M. A., Béchet J. M., Montagnier L., Hovanessian A. G. The cytopathic effect of HIV is associated with apoptosis. Virology. 1991 Dec;185(2):829–839. doi: 10.1016/0042-6822(91)90554-o. [DOI] [PubMed] [Google Scholar]
  27. Levine B., Griffin D. E. Molecular analysis of neurovirulent strains of Sindbis virus that evolve during persistent infection of scid mice. J Virol. 1993 Nov;67(11):6872–6875. doi: 10.1128/jvi.67.11.6872-6875.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Levine B., Huang Q., Isaacs J. T., Reed J. C., Griffin D. E., Hardwick J. M. Conversion of lytic to persistent alphavirus infection by the bcl-2 cellular oncogene. Nature. 1993 Feb 25;361(6414):739–742. doi: 10.1038/361739a0. [DOI] [PubMed] [Google Scholar]
  29. Lin K. I., Lee S. H., Narayanan R., Baraban J. M., Hardwick J. M., Ratan R. R. Thiol agents and Bcl-2 identify an alphavirus-induced apoptotic pathway that requires activation of the transcription factor NF-kappa B. J Cell Biol. 1995 Dec;131(5):1149–1161. doi: 10.1083/jcb.131.5.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Martin D. P., Schmidt R. E., DiStefano P. S., Lowry O. H., Carter J. G., Johnson E. M., Jr Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation. J Cell Biol. 1988 Mar;106(3):829–844. doi: 10.1083/jcb.106.3.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Meyaard L., Otto S. A., Jonker R. R., Mijnster M. J., Keet R. P., Miedema F. Programmed death of T cells in HIV-1 infection. Science. 1992 Jul 10;257(5067):217–219. doi: 10.1126/science.1352911. [DOI] [PubMed] [Google Scholar]
  32. Moench T. R., Griffin D. E. Immunocytochemical identification and quantitation of the mononuclear cells in the cerebrospinal fluid, meninges, and brain during acute viral meningoencephalitis. J Exp Med. 1984 Jan 1;159(1):77–88. doi: 10.1084/jem.159.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Morey A. L., Ferguson D. J., Fleming K. A. Ultrastructural features of fetal erythroid precursors infected with parvovirus B19 in vitro: evidence of cell death by apoptosis. J Pathol. 1993 Feb;169(2):213–220. doi: 10.1002/path.1711690207. [DOI] [PubMed] [Google Scholar]
  34. Ohno K., Okamoto Y., Miyazawa T., Mikami T., Watari T., Goitsuka R., Tsujimoto H., Hasegawa A. Induction of apoptosis in a T lymphoblastoid cell line infected with feline immunodeficiency virus. Arch Virol. 1994;135(1-2):153–158. doi: 10.1007/BF01309772. [DOI] [PubMed] [Google Scholar]
  35. Portera-Cailliau C., Hedreen J. C., Price D. L., Koliatsos V. E. Evidence for apoptotic cell death in Huntington disease and excitotoxic animal models. J Neurosci. 1995 May;15(5 Pt 2):3775–3787. doi: 10.1523/JNEUROSCI.15-05-03775.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rey-Cuille M. A., Galabru J., Laurent-Crawford A., Krust B., Montagnier L., Hovanessian A. G. HIV-2 EHO isolate has a divergent envelope gene and induces single cell killing by apoptosis. Virology. 1994 Jul;202(1):471–476. doi: 10.1006/viro.1994.1364. [DOI] [PubMed] [Google Scholar]
  37. TAYLOR R. M., HURLBUT H. S., WORK T. H., KINGSTON J. R., FROTHINGHAM T. E. Sindbis virus: a newly recognized arthropodtransmitted virus. Am J Trop Med Hyg. 1955 Sep;4(5):844–862. doi: 10.4269/ajtmh.1955.4.844. [DOI] [PubMed] [Google Scholar]
  38. Takizawa T., Matsukawa S., Higuchi Y., Nakamura S., Nakanishi Y., Fukuda R. Induction of programmed cell death (apoptosis) by influenza virus infection in tissue culture cells. J Gen Virol. 1993 Nov;74(Pt 11):2347–2355. doi: 10.1099/0022-1317-74-11-2347. [DOI] [PubMed] [Google Scholar]
  39. Tolskaya E. A., Romanova L. I., Kolesnikova M. S., Ivannikova T. A., Smirnova E. A., Raikhlin N. T., Agol V. I. Apoptosis-inducing and apoptosis-preventing functions of poliovirus. J Virol. 1995 Feb;69(2):1181–1189. doi: 10.1128/jvi.69.2.1181-1189.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tucker P. C., Griffin D. E. Mechanism of altered Sindbis virus neurovirulence associated with a single-amino-acid change in the E2 Glycoprotein. J Virol. 1991 Mar;65(3):1551–1557. doi: 10.1128/jvi.65.3.1551-1557.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tucker P. C., Strauss E. G., Kuhn R. J., Strauss J. H., Griffin D. E. Viral determinants of age-dependent virulence of Sindbis virus for mice. J Virol. 1993 Aug;67(8):4605–4610. doi: 10.1128/jvi.67.8.4605-4610.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ubol S., Griffin D. E. Identification of a putative alphavirus receptor on mouse neural cells. J Virol. 1991 Dec;65(12):6913–6921. doi: 10.1128/jvi.65.12.6913-6921.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ubol S., Tucker P. C., Griffin D. E., Hardwick J. M. Neurovirulent strains of Alphavirus induce apoptosis in bcl-2-expressing cells: role of a single amino acid change in the E2 glycoprotein. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5202–5206. doi: 10.1073/pnas.91.11.5202. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES