Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Aug;70(8):5329–5335. doi: 10.1128/jvi.70.8.5329-5335.1996

Induction of apoptosis by La Crosse virus infection and role of neuronal differentiation and human bcl-2 expression in its prevention.

A Pekosz 1, J Phillips 1, D Pleasure 1, D Merry 1, F Gonzalez-Scarano 1
PMCID: PMC190490  PMID: 8764043

Abstract

La Crosse virus causes a highly cytopathic infection in cultured cells and in the murine central nervous system (CNS), with widespread neuronal destruction. In some viral infections of the CNS, apoptosis, or programmed cell death, has been proposed as a mechanism for cytopathology (Y. Shen and T. E. Shenk, Curr. Opin. Genet. Dev. 5:105-111, 1995). To determine whether apoptosis plays a role in La Crosse virus-induced cell death, we performed experiments with newborn mice and two neural tissue culture models. Newborn mice infected with La Crosse virus showed evidence of apoptosis with the terminal deoxynucleotidyl transferase-mediated nicked-end labeling (TUNEL) assay and, concomitantly, histopathological suggestion of neuronal dropout. Infection of tissue culture cells also resulted in DNA fragmentation, TUNEL reactivity, and morphological changes in the nuclei characteristic of apoptotic cells. As in one other system (S. Ubol, P. C. Tucker, D. E. Griffin, and J. M. Hardwick, Proc. Natl. Acad. Sci. USA 91:5202-5206, 1994), expression of the human proto-oncogene bcl-2 was able to protect one neuronal cell line, N18-RE-105, from undergoing apoptosis after La Crosse virus infection and prolonged the survival of infected cells. Nevertheless, expression of bcl-2 did not prevent eventual cytopathicity. However, a human neuronal cell line, NT2N, was resistant to both apoptosis and other types of cytopathicity after infection with La Crosse virus, reaffirming the complexity of cell death. Our results show that apoptosis is an important consequence of La Crosse virus infection in vivo and in vitro.

Full Text

The Full Text of this article is available as a PDF (362.4 KB).

Selected References

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

  1. Alvarez-Buylla A., Lois C. Neuronal stem cells in the brain of adult vertebrates. Stem Cells. 1995 May;13(3):263–272. doi: 10.1002/stem.5530130307. [DOI] [PubMed] [Google Scholar]
  2. Bredesen D. E. Neural apoptosis. Ann Neurol. 1995 Dec;38(6):839–851. doi: 10.1002/ana.410380604. [DOI] [PubMed] [Google Scholar]
  3. Brooks M. A., Ali A. N., Turner P. C., Moyer R. W. A rabbitpox virus serpin gene controls host range by inhibiting apoptosis in restrictive cells. J Virol. 1995 Dec;69(12):7688–7698. doi: 10.1128/jvi.69.12.7688-7698.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bump N. J., Hackett M., Hugunin M., Seshagiri S., Brady K., Chen P., Ferenz C., Franklin S., Ghayur T., Li P. Inhibition of ICE family proteases by baculovirus antiapoptotic protein p35. Science. 1995 Sep 29;269(5232):1885–1888. doi: 10.1126/science.7569933. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cully J. F., Jr, Streit T. G., Heard P. B. Transmission of La Crosse virus by four strains of Aedes albopictus to and from the eastern chipmunk (Tamias striatus). J Am Mosq Control Assoc. 1992 Sep;8(3):237–240. [PubMed] [Google Scholar]
  7. Dickson D. W., Lee S. C., Mattiace L. A., Yen S. H., Brosnan C. Microglia and cytokines in neurological disease, with special reference to AIDS and Alzheimer's disease. Glia. 1993 Jan;7(1):75–83. doi: 10.1002/glia.440070113. [DOI] [PubMed] [Google Scholar]
  8. Eastman C. L., Guilarte T. R., Lever J. R. Uptake of 3-hydroxykynurenine measured in rat brain slices and in a neuronal cell line. Brain Res. 1992 Jul 3;584(1-2):110–116. doi: 10.1016/0006-8993(92)90883-b. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Geiger K. D., Gurushanthaiah D., Howes E. L., Lewandowski G. A., Reed J. C., Bloom F. E., Sarvetnick N. E. Cytokine-mediated survival from lethal herpes simplex virus infection: role of programmed neuronal death. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3411–3415. doi: 10.1073/pnas.92.8.3411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Griot C., Pekosz A., Davidson R., Stillmock K., Hoek M., Lukac D., Schmeidler D., Cobbinah I., Gonzalez-Scarano F., Nathanson N. Replication in cultured C2C12 muscle cells correlates with the neuroinvasiveness of California serogroup bunyaviruses. Virology. 1994 Jun;201(2):399–403. doi: 10.1006/viro.1994.1308. [DOI] [PubMed] [Google Scholar]
  12. Griot C., Pekosz A., Lukac D., Scherer S. S., Stillmock K., Schmeidler D., Endres M. J., Gonzalez-Scarano F., Nathanson N. Polygenic control of neuroinvasiveness in California serogroup bunyaviruses. J Virol. 1993 Jul;67(7):3861–3867. doi: 10.1128/jvi.67.7.3861-3867.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hardy M., Younkin D., Tang C. M., Pleasure J., Shi Q. Y., Williams M., Pleasure D. Expression of non-NMDA glutamate receptor channel genes by clonal human neurons. J Neurochem. 1994 Aug;63(2):482–489. doi: 10.1046/j.1471-4159.1994.63020482.x. [DOI] [PubMed] [Google Scholar]
  14. Hay B. A., Wassarman D. A., Rubin G. M. Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death. Cell. 1995 Dec 29;83(7):1253–1262. doi: 10.1016/0092-8674(95)90150-7. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Hockenbery D., Nuñez G., Milliman C., Schreiber R. D., Korsmeyer S. J. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990 Nov 22;348(6299):334–336. doi: 10.1038/348334a0. [DOI] [PubMed] [Google Scholar]
  17. Janssen R. S., Nathanson N., Endres M. J., Gonzalez-Scarano F. Virulence of La Crosse virus is under polygenic control. J Virol. 1986 Jul;59(1):1–7. doi: 10.1128/jvi.59.1.1-7.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Janssen R., Gonzalez-Scarano F., Nathanson N. Mechanisms of bunyavirus virulence. Comparative pathogenesis of a virulent strain of La Crosse and an avirulent strain of Tahyna virus. Lab Invest. 1984 Apr;50(4):447–455. [PubMed] [Google Scholar]
  19. Kalfayan B. Pathology of La Crosse virus infection in humans. Prog Clin Biol Res. 1983;123:179–186. [PubMed] [Google Scholar]
  20. Levine B., Hardwick J. M., Trapp B. D., Crawford T. O., Bollinger R. C., Griffin D. E. Antibody-mediated clearance of alphavirus infection from neurons. Science. 1991 Nov 8;254(5033):856–860. doi: 10.1126/science.1658936. [DOI] [PubMed] [Google Scholar]
  21. Lewis J., Wesselingh S. L., Griffin D. E., Hardwick J. M. Alphavirus-induced apoptosis in mouse brains correlates with neurovirulence. J Virol. 1996 Mar;70(3):1828–1835. doi: 10.1128/jvi.70.3.1828-1835.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Llanes C., Collman R. G., Hrin R., Kolson D. L. Acetylcholinesterase expression in NTera 2 human neuronal cells: a model for developmental expression in the nervous system. J Neurosci Res. 1995 Dec 15;42(6):791–802. doi: 10.1002/jnr.490420608. [DOI] [PubMed] [Google Scholar]
  24. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  25. Murphy T. H., De Long M. J., Coyle J. T. Enhanced NAD(P)H:quinone reductase activity prevents glutamate toxicity produced by oxidative stress. J Neurochem. 1991 Mar;56(3):990–995. doi: 10.1111/j.1471-4159.1991.tb02019.x. [DOI] [PubMed] [Google Scholar]
  26. Murphy T. H., Schnaar R. L., Coyle J. T., Sastre A. Glutamate cytotoxicity in a neuronal cell line is blocked by membrane depolarization. Brain Res. 1988 Sep 13;460(1):155–160. doi: 10.1016/0006-8993(88)91216-4. [DOI] [PubMed] [Google Scholar]
  27. Olsen C. W., Kehren J. C., Dybdahl-Sissoko N. R., Hinshaw V. S. bcl-2 alters influenza virus yield, spread, and hemagglutinin glycosylation. J Virol. 1996 Jan;70(1):663–666. doi: 10.1128/jvi.70.1.663-666.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pekosz A., Griot C., Nathanson N., Gonzalez-Scarano F. Tropism of bunyaviruses: evidence for a G1 glycoprotein-mediated entry pathway common to the California serogroup. Virology. 1995 Dec 20;214(2):339–348. doi: 10.1006/viro.1995.0043. [DOI] [PubMed] [Google Scholar]
  29. Pekosz A., Griot C., Stillmock K., Nathanson N., Gonzalez-Scarano F. Protection from La Crosse virus encephalitis with recombinant glycoproteins: role of neutralizing anti-G1 antibodies. J Virol. 1995 Jun;69(6):3475–3481. doi: 10.1128/jvi.69.6.3475-3481.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shen Y., Shenk T. E. Viruses and apoptosis. Curr Opin Genet Dev. 1995 Feb;5(1):105–111. doi: 10.1016/s0959-437x(95)90061-6. [DOI] [PubMed] [Google Scholar]
  31. Stinski M. F., Roehr T. J. Activation of the major immediate early gene of human cytomegalovirus by cis-acting elements in the promoter-regulatory sequence and by virus-specific trans-acting components. J Virol. 1985 Aug;55(2):431–441. doi: 10.1128/jvi.55.2.431-441.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Subramanian T., Tarodi B., Chinnadurai G. p53-independent apoptotic and necrotic cell deaths induced by adenovirus infection: suppression by E1B 19K and Bcl-2 proteins. Cell Growth Differ. 1995 Feb;6(2):131–137. [PubMed] [Google Scholar]
  33. Tarodi B., Subramanian T., Chinnadurai G. Epstein-Barr virus BHRF1 protein protects against cell death induced by DNA-damaging agents and heterologous viral infection. Virology. 1994 Jun;201(2):404–407. doi: 10.1006/viro.1994.1309. [DOI] [PubMed] [Google Scholar]
  34. Tyler K. L., Squier M. K., Rodgers S. E., Schneider B. E., Oberhaus S. M., Grdina T. A., Cohen J. J., Dermody T. S. Differences in the capacity of reovirus strains to induce apoptosis are determined by the viral attachment protein sigma 1. J Virol. 1995 Nov;69(11):6972–6979. doi: 10.1128/jvi.69.11.6972-6979.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Westmoreland S. V., Kolson D., González-Scarano F. Toxicity of TNF alpha and platelet activating factor for human NT2N neurons: a tissue culture model for human immunodeficiency virus dementia. J Neurovirol. 1996 Apr;2(2):118–126. doi: 10.3109/13550289609146545. [DOI] [PubMed] [Google Scholar]
  37. Wyllie A. H. The genetic regulation of apoptosis. Curr Opin Genet Dev. 1995 Feb;5(1):97–104. doi: 10.1016/s0959-437x(95)90060-8. [DOI] [PubMed] [Google Scholar]
  38. Yasuda T., Grinspan J., Stern J., Franceschini B., Bannerman P., Pleasure D. Apoptosis occurs in the oligodendroglial lineage, and is prevented by basic fibroblast growth factor. J Neurosci Res. 1995 Feb 15;40(3):306–317. doi: 10.1002/jnr.490400304. [DOI] [PubMed] [Google Scholar]
  39. Younkin D. P., Tang C. M., Hardy M., Reddy U. R., Shi Q. Y., Pleasure S. J., Lee V. M., Pleasure D. Inducible expression of neuronal glutamate receptor channels in the NT2 human cell line. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2174–2178. doi: 10.1073/pnas.90.6.2174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Zhu H., Shen Y., Shenk T. Human cytomegalovirus IE1 and IE2 proteins block apoptosis. J Virol. 1995 Dec;69(12):7960–7970. doi: 10.1128/jvi.69.12.7960-7970.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES