미생물학회지 (Korean Journal of Microbiology)

  • 제38권3호
  • /
  • Pages.221-229
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  • 2002
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

지속감염세포에서 분리된 일본뇌염바이러스 Plaque Morphology Mutants의 복제 및 감염특성

Replication and Pathogenesis of Plaque Morphology Mutants Derived from Vero Cells with Japanese Encephalitis Virus Persistency

  • 윤성욱 (경희대학교 생물학과 및 기초과학연구소) ;
  • 정용석 (경희대학교 생물학과 및 기초과학연구소)
  • 발행 : 2002.09.01
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초록

일본뇌염바이러스(Japanese encephalitis virus, JEV) Nakayama strain을 초기 multiplicity of infection 5.0으로 Vero세포에 감염하여 1년 이상 안정적으로 바이러스를 방출하는 지속감염(persistently-infected)세포주를 확립하였다. 지속감염 세포에서 지속적으로 방출되는 총 11 개의 Plaque 형태 변이바이러스(morphology mutants)클론을 확보하였다. 분리된 변이바이러스의 복제효율을 분석한 결과 생물학적 표현형과 복제효율은 유의하게 상관하였다. 변이바이러스 RNA 게놈 양 말단의 non-coding region 및 envelop 단백질의 ORF에서는 유의한 염기서열 변화가 관찰되지 않아 JEV 약독화에 새로운 인자가 추가로 관여할 가능성을 제시하였다. 변이바이러스에 감염된 신선한 Vero세포는 wild-type JEV의 일반적 감염성상과 다르게 대다수의 세포가 유의할 만한 세포병변현상을 나타내지 않았다. 감염된 Vero세포에서 wild-type JEV 및 large plaque을 형성하는 변이바이러스의 경우 mRNA와 함께 Bcl-2의 발현은 모두 유의하게 감소하였으며 p53은 뚜렷하게 증가하였다. 반면 small plaque을 형성하는 변이바이러스의 감염세포에서는 Bcl-2와 p53 모두 유의한 변화를 볼 수 없었다. 이상의 결과들과 함께, 감염된 Vero세포의 internucleosomal DNA fragmentation과DNA profile의 유형분석에 따르면 궁극적 인 세포병변효과의 변화는 변이바이러스의 복제효율과 더불어 p53에 비의존적인 apoptosis 수위의 전반적인 감소에 기인하는 것으로 판단된다.

Japanese encephalitis virus (JEV) persistence was established and maintained in Vero cell culture for over 1 year. Eleven clones of plaque morphology mutant JEV, with large and small plaque sizes, were obtained from the cell culture supernatant. Genomic RNA replication efficiency of the mutants in naive Vero cell appeared to correspond to their different plaque sizes. No significant changes in envelop protein ORF or in non-coding regions at both ends of the RNA genome suggested that there could be an unidentified factor(s) playing role in JEV attenuation. Unlike to the replication of wild-type JEV, the mutants did not induce severe degree of cytopathic effect in Vero cells upon infection. While obvious decrease of Bcl-2 and its mRNA expression and sharp increase of p53 in naive Vero cells infected with either wild-type JEV or the large plaque-forming mutant, those changes were not observed with the small plaque-forming one. Together with these observation, internucleosomal DNA fragmentation and chromosomal DNA profile in the Vero cells infected with the mutants suggest that an overall changes in cytopathic effect in the plaque morphology mutants-infected cells should be primarily due to the reduced genomic RNA replication and the compromised degree of p53-independent apoptosis by the virus infection at least in part.

키워드

참고문헌

  1. Virology v.207 Growth restricted dengue virus mutants containing deletions in the 5' noncoding region of the RNA genome. Cahour, A.;A. Pletnev;M. Vazeille-Falcoz;L. Rosen;C-J. Lai https://doi.org/10.1006/viro.1995.1052
  2. J. Virol. v.71 RNA-protein interactions: involvement of NS3, NS5 and 3' noncoding regions of Japanese encephalitis virus genomic RNA Chen, C-J.;M-D. Kuo;L-J. Chien;S-L. Hsu;Y-M. Wang;J-H. Lin
  3. Virology v.217 Persistence of Japanese encephalitis virus is associated with abnormal expression of the nonstructural protein NS1 in host cells Chen, L-K.;C-L. Liao.;C-G. Lin;S-C. Lai;C-I. Liu;S-H. Ma;Y-Y. Huang;Y-L. Lin https://doi.org/10.1006/viro.1996.0109
  4. Virology v.223 Generation and characterization of organ-tropism mutants of Japanese encephalitis virus in vivo and in vitro Chen,L-K.;Y-L. Lin;C-L. Liao.;C-G. Lin.;Y-L Huang.;C-T. Yeh.;S-C. Jan;C. Chin https://doi.org/10.1006/viro.1996.0457
  5. Virology v.255 Rubella virus-induced apoptosis varies among cell lines and is modulated by Bcl-XL and caspase inhibitors Duncan, R.;J. Muller;N. Lee;A. Esmaili;H.L. Nakhasi https://doi.org/10.1006/viro.1998.9562
  6. Vaccine v.12 Comparison of structural proteins among antigenically different Japanese encephalitis virus strains Hasegawa, H.;M. Yoshida;S. Fujita;Y. Kobayashi https://doi.org/10.1016/0264-410X(94)90294-1
  7. Virology v.264 Apoptosis is induced by hantaviruses in cultured cells Kang, J-I;S-H. Park;P-W. Lee;B-Y. Ahn https://doi.org/10.1006/viro.1999.9896
  8. J. Virol. v.68 Completion of Kunjin virus RNA sequence and recovery of an infectious RNA transcribed from stably cloned full-length cDNA Khromykh, A.A.;E.G. Westaway
  9. Vaccines 92. Modern Approaches to New Vaccines Including Prevention of AIDS Infectious RNA transcribed from stably cloned dengue virus cDNA: consruction of growth restricted dengue virus mutants Lai, CJ.;R. Men;M. Pethel;M. Bray;H.S. Ginsburg(ed.);F. Brown(ed.);R.M. Chanock(ed.);R.A. Lerner(ed.)
  10. J. Virol. v.71 Effect of enforced expression of huma bcl-2 on Japanese encephalitis virus-induced apoptosis in cultured cells Liao, C-L.;Y. Lin;J-J. Wang;Y-L. Huang;C-T. Yeh;S-H. Ma;L-K. Chen
  11. J. Virol. v.72 Antiapoptotic but antiviral function of human bcl-2 assists establishment of Japanese encephalitis virus persistence in cultured cells Liao, C-L.;Y-L. Lin;S-C. Shen;J-Y. Shen;H-L. Su;Y-L. Huang;S-H. Ma;Y-C. Sun;K-P. Chen;L-K. Chen
  12. Virology v.173 Characterization of Japanese encephalitis virus envelope protein expressed by recombinant baculoviruses Matsuura Y.;M. Miyamoto;T. Sato;C. Morita;K. Yasui https://doi.org/10.1016/0042-6822(89)90580-1
  13. Virology v.259 Involvement of a p53-dependent pathway in Rubella virus-induced apoptosis Megyeri, K.;K. Berencsi;T.D. Halazonetis;G.C. Prendergast;G. Gri;S.A. Plotkin;G. Rovera;E. Gonczol https://doi.org/10.1006/viro.1999.9757
  14. J. Virol. v.70 Dengue type 4 virus mutants containing deletions in the 3'noncoding region of the RNA genome: analysis of growth restriction in cell culture and altered viremia pattern and immunogenicity Men, R.;M. Bray;D. Clark;R.M. Chanock;C-J. Lai
  15. J. Virol. v.71 Genetic analysis of the yellow fever virus NS1 protein: identification of a temperaturesensitive mutation which blocks RNA accumulation Muylaert, I.R.;R. Galler;C.M. Rice
  16. SA-14-14-2. Virology v.177 Nucleotide sequence of the virulent SA-14 strain of Japanese encephalitis virus and its attenuated vaccine derivative Nitayaphan, S.;J.A;G-J.J. Chang;D.W. Trent
  17. Virology v.218 Molecular characterization of the Japanese encephalitis serocomplex of the flavivirus genus Poidinger, M.;R.A. Hall;J.S. Mackenzie https://doi.org/10.1006/viro.1996.0213
  18. J. Med. Virol. v.40 Persistence of Japanese encephalitis virus in the human nervous system Ravi, V.;A.S. Desai;P.K. Shenoy;P. Satishchandra;A. Chandramuki;M. Gourie-Devi https://doi.org/10.1002/jmv.1890400412
  19. J. Virol. v.67 Genetic studies of flavivirus resistance in inbred strains derived from wild mice: evidence for a new resistant allele at the flavivirus resistance locus(flv) Sangster, M.Y.;D.B. Heliams;J.S. Mackenzie;G.R. Shellam
  20. J. Virol. v.71 bcl-2 acts early to restrict Semliki forest virus replication and delay virus -induced programmed cell death Scallan, M.F.;T.E. Allsopp;J.K. Fazakerley
  21. J. Virol. v.24 Persistent infection of cultured mammalian cells by Japanese encephalitis virus Schmaljohn, C.S.;C.D. Blair
  22. J. Virol. v.31 Clonal analysis of mammalian cell cultures persistently infected with Japanese encephalitis virus Schmaljohn, C.S.;C.D. Blair
  23. Virology v.89 The generation and propagation of defective-interfering particles of Semliki forest virus in different cell types Stark, C.;S.I.T. Kennedy https://doi.org/10.1016/0042-6822(78)90060-0
  24. Virology v.161 Complete nucleotide sequence of the Japanese encephalitis virus genome RNA Sumiyoshi, H.;C. Mori;I. Fuke;K. Morita;S. Kuhara;J. Kondou;Y. Kikuchi;H. Nagamatu;A. Igarashi https://doi.org/10.1016/0042-6822(87)90144-9
  25. J. Infect. Dis. v.171 Charaterization of a highly attenuated Japanese encephalitis virus generated from molecularly cloned cDNA Sumiyoshi, H.;G.H. Tignor;R. E. Hope https://doi.org/10.1093/infdis/171.5.1144