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Current IssueEditorial BoardOnline First ArticlesArchive

volume 49 issue 5 (october 2015) : 612-617,   Doi: 10.18805/ijar.5572

Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro

H
H. Prabhavathy
K
K.M. Palanivel
1Department of Veterinary Epidemiology and Preventive Medicine, Madras Veterinary College, Chennai-600 007, India.
Cite article:- Prabhavathy H., Palanivel K.M. (2025). Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro. Indian Journal of Animal Research. 49(5): 612-617. doi: 10.18805/ijar.5572.

ABSTRACT

An attempt had been made in this study to know the ability of Indian street rabies virus isolates to induce apoptosis in murine neuroblastoma cells. Thirty Indian street virus isolates were tested by fluorescent antibody technique (FAT) among which 20 samples was positive. These positive samples were used to infect murine neuroblastoma (MNA) cells. Four street rabies virus isolates and challenge virus strain (CVS) were used in this study to show their ability to induce apoptosis. Total RNA was isolated and cDNA was synthesized by using gene specific primers pertaining to Caspase-1, BAD (Proapoptotic gene) and Glycoprotein gene of rabies virus. Challenge Virus Standard has been used as control in the amplification as well as in the demonstration of apoptosis. The expression of Caspase-1, BAD and glycoprotein genes in the murine neuroblastoma cells in comparision with CVS was measured by using densitometer. Street virus isolates expressed higher levels of Caspase-1 and BAD in comparison with CVS. Further TUNEL staining method showed higher apoptotic index when compared to CVS. This study is an initial attempt to exhibit the ability of Indian rabies street virus isolates to induce apoptosis in murine neuroblastoma cells (in vitro).

REFERENCES

  1. Alcami, A., Koszinowski.,U.H.(2000). Viral mechanisms of immune evasion. Trends Microbial,8: 410-418
  2. Amoult, D., Parone, P.,Martinou, J.C., Antonss, B., Estaquier J and Ameisen, J.C. (2002). Mitochondrial release of apoptosis- inducing factor occurs downstream ofcytochrome-C releasein response to several Pro-apoptotic stimuli. J.CellBiol,159: 923-929.
  3. Choi, C., and Benveniste, E.N.(2004). Fas Ligand/Fas System in the brain: regulator of immune and apoptotic responses. Brain Res Rev,44: 65-81.
  4. Dean, D.J.and Abelseth, M.K.(1973). The fluroscent antibody test. In: Laboratory techniques in rabies 3rd Eds., M.M.Kaplan and H.Koprowski.WHO, Geneva: pp 73-84.
  5. Galelli A., BaloulL.andLafon, M.(2000). Abortive rabies virus central nervous infection is controlled by T.Lymphocyte local recruitment and induction of apoptosis. J Neurovirol, 6: 359-372.
  6. Harada, K.H and Grant. (2003). Apoptosis regulators. Rev.Clinic, Exp. Hematol,7:117-138.
  7. Hon Kit Wong, F., Michael, W., Andreas, V.,Ewa, M., Andreas S. andTolkovsky, M.(1994).Mutually Exclusive Subsets ofBH3- only proteins are activated by the p53 and c-junN-Terminal Kinase/c-junsignaling pathways during cortical Neuron Apoptosisinduced by arsenite. Molecular and Cellular Biology,25: 8732-8747.
  8. Iwasaki, Y., and Tobita, M.(2002). Rabies virus: In Jackson, A.C. andWunner,W.H (eds)., Academic press, san Diego. Pathology, pp: 283-306.
  9. Morimoto, K.D., Hooper,C., Spitsin,S., KoprowskiH. and Dietzschold, B.(1999).Pathogenecity of different rabies virus variants inversely correlates with apoptosis and rabies virus glycoprotein expression in infected primary neuron cultures. J.Virol, 73.510-518.
  10. Morimoto, K.H., Foley,D., McGettigan,J.P., Schnell,M.J. andDietzschold, B.(2000). Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetic approach. J.Neurovirol, 6. 373-381.
  11. Palanivel,K.M., Saravanan,S., Selvaraju,G., Geetha,M and Jayakumar, R. (2011). Acase of furious rabies in a Man following unusual long incubation period. J Vet Pub Hlth,9:127-128.
  12. Reid, J.E., andJackson, A.C.(2001). Experimental rabies virus infection in Artibeusjamaicensis bat with CVS-24 variants. J. NeuroVirol, 7: 511-517.
  13. Schnell, M.J.,Tan,G.S and.Dietzschold, B.(2005). The application of reverse geneticstechnology in the study of rabies virus pathogenesis and for the development of novelRV vaccines. J.Neurovirol, 11:76-81.
  14. Smith,J.S andKing, A.K.(1996). Monoclonal antibodies for identification of rabies and non-rabies lyssa viruses. In: Laboratory Techniques in Rabies.(Meslin, F.X.,Kaplan,M.M.and.Koprowski, H et al): 4thedn. World Health Organization, Geneva,pp 145-155.
  15. Theerasurakan,S. and Ubol, S.(1998). Apoptosis induction in brain during the fixed strain ofrabies virus infection correlates with onset and severity of illness. J.Neurovirol,4:407-414.
  16. Tirumurugaan,K.G. (2001). Expression of glycoprotein of rabies virus in eukaryotic cells. Ph.D.,Thesis submitted to the Tamilnadu University of Veterinary and animal Sciences.Ubol, S., Kasisith, J.,Pitidhammabhorn, D. and Tepsum ethanolV. (2005). Screening of pro-apoptotic genes up regulated in an experimental street rabies virus infectedneonatal mares brain. Microbiloimmunol, 49(5): 423-431.
  17. Wesselingh,S.L.,Levine, B., Fox, R.J., Choi SandGriffin, D.E.(1994). Intra cerebral cytokine MRNA expression during fatal and nonfatal alphavirus encephalitissuggests a predomoinant type 2T cell response. J.Immunol,152: 1289-1297.
  18. World Health Organization. (1994). Terrestrial Manual, pp: 145-155.
  19. Zhang, W.H., Wang, X., Narayaman, M., Zhang, Y., Huo,C., Reed J.C. and Friedlander, R.M.(2003). Fundamental role of the Rip2/ caspase-1 pathway in hypoxia andischemia-induced neuronal cell death. Proc. Natl.Acad.Sc, U.S.A.100: 16012-16017.
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Indian Journal of Animal Research
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    volume 49 issue 5 (october 2015) : 612-617,   Doi: 10.18805/ijar.5572

    Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro

    H
    H. Prabhavathy
    K
    K.M. Palanivel
    1Department of Veterinary Epidemiology and Preventive Medicine, Madras Veterinary College, Chennai-600 007, India.
    Cite article:- Prabhavathy H., Palanivel K.M. (2025). Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro. Indian Journal of Animal Research. 49(5): 612-617. doi: 10.18805/ijar.5572.

    ABSTRACT

    An attempt had been made in this study to know the ability of Indian street rabies virus isolates to induce apoptosis in murine neuroblastoma cells. Thirty Indian street virus isolates were tested by fluorescent antibody technique (FAT) among which 20 samples was positive. These positive samples were used to infect murine neuroblastoma (MNA) cells. Four street rabies virus isolates and challenge virus strain (CVS) were used in this study to show their ability to induce apoptosis. Total RNA was isolated and cDNA was synthesized by using gene specific primers pertaining to Caspase-1, BAD (Proapoptotic gene) and Glycoprotein gene of rabies virus. Challenge Virus Standard has been used as control in the amplification as well as in the demonstration of apoptosis. The expression of Caspase-1, BAD and glycoprotein genes in the murine neuroblastoma cells in comparision with CVS was measured by using densitometer. Street virus isolates expressed higher levels of Caspase-1 and BAD in comparison with CVS. Further TUNEL staining method showed higher apoptotic index when compared to CVS. This study is an initial attempt to exhibit the ability of Indian rabies street virus isolates to induce apoptosis in murine neuroblastoma cells (in vitro).

    REFERENCES

    1. Alcami, A., Koszinowski.,U.H.(2000). Viral mechanisms of immune evasion. Trends Microbial,8: 410-418
    2. Amoult, D., Parone, P.,Martinou, J.C., Antonss, B., Estaquier J and Ameisen, J.C. (2002). Mitochondrial release of apoptosis- inducing factor occurs downstream ofcytochrome-C releasein response to several Pro-apoptotic stimuli. J.CellBiol,159: 923-929.
    3. Choi, C., and Benveniste, E.N.(2004). Fas Ligand/Fas System in the brain: regulator of immune and apoptotic responses. Brain Res Rev,44: 65-81.
    4. Dean, D.J.and Abelseth, M.K.(1973). The fluroscent antibody test. In: Laboratory techniques in rabies 3rd Eds., M.M.Kaplan and H.Koprowski.WHO, Geneva: pp 73-84.
    5. Galelli A., BaloulL.andLafon, M.(2000). Abortive rabies virus central nervous infection is controlled by T.Lymphocyte local recruitment and induction of apoptosis. J Neurovirol, 6: 359-372.
    6. Harada, K.H and Grant. (2003). Apoptosis regulators. Rev.Clinic, Exp. Hematol,7:117-138.
    7. Hon Kit Wong, F., Michael, W., Andreas, V.,Ewa, M., Andreas S. andTolkovsky, M.(1994).Mutually Exclusive Subsets ofBH3- only proteins are activated by the p53 and c-junN-Terminal Kinase/c-junsignaling pathways during cortical Neuron Apoptosisinduced by arsenite. Molecular and Cellular Biology,25: 8732-8747.
    8. Iwasaki, Y., and Tobita, M.(2002). Rabies virus: In Jackson, A.C. andWunner,W.H (eds)., Academic press, san Diego. Pathology, pp: 283-306.
    9. Morimoto, K.D., Hooper,C., Spitsin,S., KoprowskiH. and Dietzschold, B.(1999).Pathogenecity of different rabies virus variants inversely correlates with apoptosis and rabies virus glycoprotein expression in infected primary neuron cultures. J.Virol, 73.510-518.
    10. Morimoto, K.H., Foley,D., McGettigan,J.P., Schnell,M.J. andDietzschold, B.(2000). Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetic approach. J.Neurovirol, 6. 373-381.
    11. Palanivel,K.M., Saravanan,S., Selvaraju,G., Geetha,M and Jayakumar, R. (2011). Acase of furious rabies in a Man following unusual long incubation period. J Vet Pub Hlth,9:127-128.
    12. Reid, J.E., andJackson, A.C.(2001). Experimental rabies virus infection in Artibeusjamaicensis bat with CVS-24 variants. J. NeuroVirol, 7: 511-517.
    13. Schnell, M.J.,Tan,G.S and.Dietzschold, B.(2005). The application of reverse geneticstechnology in the study of rabies virus pathogenesis and for the development of novelRV vaccines. J.Neurovirol, 11:76-81.
    14. Smith,J.S andKing, A.K.(1996). Monoclonal antibodies for identification of rabies and non-rabies lyssa viruses. In: Laboratory Techniques in Rabies.(Meslin, F.X.,Kaplan,M.M.and.Koprowski, H et al): 4thedn. World Health Organization, Geneva,pp 145-155.
    15. Theerasurakan,S. and Ubol, S.(1998). Apoptosis induction in brain during the fixed strain ofrabies virus infection correlates with onset and severity of illness. J.Neurovirol,4:407-414.
    16. Tirumurugaan,K.G. (2001). Expression of glycoprotein of rabies virus in eukaryotic cells. Ph.D.,Thesis submitted to the Tamilnadu University of Veterinary and animal Sciences.Ubol, S., Kasisith, J.,Pitidhammabhorn, D. and Tepsum ethanolV. (2005). Screening of pro-apoptotic genes up regulated in an experimental street rabies virus infectedneonatal mares brain. Microbiloimmunol, 49(5): 423-431.
    17. Wesselingh,S.L.,Levine, B., Fox, R.J., Choi SandGriffin, D.E.(1994). Intra cerebral cytokine MRNA expression during fatal and nonfatal alphavirus encephalitissuggests a predomoinant type 2T cell response. J.Immunol,152: 1289-1297.
    18. World Health Organization. (1994). Terrestrial Manual, pp: 145-155.
    19. Zhang, W.H., Wang, X., Narayaman, M., Zhang, Y., Huo,C., Reed J.C. and Friedlander, R.M.(2003). Fundamental role of the Rip2/ caspase-1 pathway in hypoxia andischemia-induced neuronal cell death. Proc. Natl.Acad.Sc, U.S.A.100: 16012-16017.
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