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Analysis of persistent infection mechanism of RNA virus

Research Project

Project/Area Number 08457101
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionThe Tokyo Metropolitan Institute of Medical Science

Principal Investigator

KOHARA Michinori  The Tokyo Metropolitan Institute of Medical Science, Department of Radiation Research, Head, 放射線医学研究部門, 研究員 (10250218)

Co-Investigator(Kenkyū-buntansha) KOHARA Kyoko  The Tokyo Metropolitan Institute of Medical Science, Department of Radiation Res, 放射線医学研究部門, 研究員 (20225478)
WAKITA Takaji  The Tokyo Metropolitan Institute of Medical Science, Department of Radiation Res, 放射線医学研究部門, 研究員 (40280789)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 1997: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1996: ¥3,900,000 (Direct Cost: ¥3,900,000)
KeywordsHepatitis C virus / HCV cDNA / Infectious clone / Immuno-electron microscopic study / HCV like-particle / 感染性cDNAクローン / 遺伝子複製
Research Abstract

One of the major impediments to the structural analysis of the HCV genome and genetic analysis of viral replication has been lack of a reliable cell culture system permissive for HCV replication. Therefore, we established the HCV replication system by using entire HCV cDNA and T7 RNA polymerase recombinant adeno virus system.
Methods : the putative full-length HCV genome (1-9603) was transiently expressed by the adeno virus/T7 RNA polymerase hybrid expression system. We transfected to a HCV high sensitive cell line (IMY cell) with HCV entire cDNA clone and infected recombinant adeno virus. The HCV particle was characterized by immuno-electron microscopy. Results : The transfected HCV genome replicated in cells, as evidenced by appearance of progeny HCV RNA and detection of negative-strand viral RNA.Immuno-electron microscopic studies have shown that the HCV like-particle was detected in the transfected cells and culture medium. We show the replication process of HCV in the cell and clarified the structure of viral particle. Conclusion : The expression system using HCV entire cDNA clone and showed the importance of 3'X region for HCV replication in the cell. The elctron microscopy study revealed the dynamics of viral core particle in the cells. Moreover the core particle was proved to fold the hexagonal structure.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Barba, G.,: "Hepatitis C Virus Core Protein shows a Cytoplasmic Localization and Associates to Cellular Lipid Storage Droplets." Proc.Natl.Acad.Sci.USA.94. 1200-1205 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kamoshita, N.: "Genetic analysis of internal ribosomal entry site on hepatitis C-virus RNA:implication for involvement of the highly-ordered structure and cell type specific trans-acting factors." Virology. 233. 9-18 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tanaka, T.: "Acute hepatitis caused by sexual or household transmission of GBV-C." J.Hepatology. 27. 1110-1112 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Akiyama, H.: "Effects of cyclosporin A on hepatitis C virus infection in bone marrow transplantation patients." Bone Marrow Transplant. 20. 993-995 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakita, T.: "Effects conditional transgene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system." J.Biol.Chem.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 小原 道法: "Modem Physician" C型肝炎とG型肝炎検査法の進歩, 5 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 小原 道法: "南江堂" C型肝癌発生とウイルス増殖, 4 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yohko K.Shimizu, Stephen M.Feinstone, Michinori Kohara, Robert H.Purcell, and Hiroshi Yoshikura: "Hepatitis C Virus : Detection mof Interacellular Virus Particles by Electron Microscopy" Hepatology. 23. 205-209 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tomiko Kashiwakuma, Akira Hasegawa, Tadahiro Kajita, Atumi Tanaka, Hiyoyuki Mori, Yohsuke Ohta, Eiji Tanaka, Kendo Kiyosawa, Takeshi Tanaka, Satoshi Tanaka, Nobu Hattori and Michinori Kohara: "Detection of hepatitis C virus specific core protein in serum of patients by a sensitive fluorescence enzyme immunoassay (FEIA). J.Immunological Methods" J.Immunological Methods. 190. 79-89 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] E.Orito, M.Mizokami, T.Tanaka, J.Y-N.Lau, K.Suzuki, M.Yamauchi, Y.Ohta, A.Hasegawa, S.Taanaka and M.Kohara: "Quantification of serum hepatitis C virus core protein level in patients chronically infected with different hepatitis C virus genotypes" GUT. 39. 876-880 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kyoko Tsukiyama-Kohara, Silvia M.Vidal, Anne-Claude Gingras, Thomas W.Glover, Samir M.Hanash, Henry Heng, and Nahum Sonenberg: "Tissue Distribution, Genomic Structure, and Chromosome Mapping of Mouse and Human Eukaryotic Initiation Factor 4E-Binding Proteins 1 and 2." Genomics. 38. 353-363 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] G.Barba, F.Harper, T.Harada, M.Kohara, S.Goulinet, Y.Matsuura, G.Eder, Zs.Schaff, M.J.Chapman, T.Miyamura, and C.Brechot.: "Hepatitis C Virus Core Protein shows a Cytoplasmic Localization and Associates to Cellular Lipid Storage Droplets." Proc.Natl.Acad.Sci.USA. 94. 1200-1205 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tokushige, K., Moradpour, D., Wakita, T., Geissler, M., Hayashi, N., Wand, JR.: "Comparison between cytomegalovirus promoter and elongation factor-1 alpha promoter-driven constructs in the establishment of cell lines expressing hepatitis C virus core protein" J.of Virological Methods. 64 (1). 73-80 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nobuhiko Kamoshita, Kyoko Tsukiyama-Kohara, Michinori Kohara, and Akio Nomoto.: "Genetic analysis of internal ribosomal entry site on hepatitis C virus RNA : implication for involvement of the highly-ordered structure and cell type specific trans-acting factors." Virology. 233. 9-18 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takeshi Tanaka, Tomoko Takeuchi, Kazuaki Inoue, Satoshi Tanaka, and Michinori Kohara: "Acute hepatitis caused by sexual or household transmission of GBV-C." J.Hepatology. 27. 1110-1112 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hideki Akiyama, Haruhiko Yoshinaga, Takeshi Tanaka, Kiyoshi Hiruma, Shu Tanikawa, Hisashi Sakamaki, Yasusuke Onozawa, Takaji Wakita, Michinori Kohara, and Bone Marrow Transplantation Team.: "Effects of cyclosporin A on hepatitis C virus infection in bone marrow transplantation patients." Bone Marrow Transplant. 20. 993-995 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takaji Wakita, Choji Taya, Asao Katsume, Junko Kato, Hiromichi Yonekawa, Yumi Kanegae, Izumu Saito, Yukiko Hayasi, Morio Koike, Michinori Kohara: "Efficient conditional trasngene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system." J.Biol.Chem.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Barba,G.,: "Hepatitis C Virus Core Protein shows a Cytoplasmic Localization and Associates to Cellular Lipid Storage Droplets." Proc.Natl.Acad.Sci.USA.94. 1200-1205 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kamoshita,N.: "Genetic analysis of internal ribosomal entry site on hepatitis C virus RNA:implication for involvement of the highly-ordered structure and cell type specific trans-acting factors." Virology. 233. 9-18 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tanaka,T.: "Acute hepatitis caused by sexual or household transmission of GBV-C." J.Hepatology. 27. 1110-1112 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Akiyama,H.: "Effects of cyclosporin A on hepatitis C virus infection in bone marrow transplantation patients." Bone Marrow Transplant. 20. 993-995 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Wakita,T.: "Efficient conditional transgene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system." J.Biol.Chem.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondo,S.: "Inhibition of telomerase increases the susceptibility of human malignant glioma cells to cisplatin-induced apoptosis." Oncogene. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kase,R.: "Immunohistochemical characterization of transgenic mice highly expressing human lysosomal α-galactosidase" Biochimica et Biophvsica Acta. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondo,T.: "Identification and chacterization of nucleophosmin/B23/numatrin which binds the anti-oncogenic transcription factor IRF-1 and manifests oncogenic activity." Oncogene. 15. 1275-1281 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Taniguchi,T.: "IRF-1:the transcription factor linking the interferon response and oncogenesis." Biochimica et Biophysica Acta. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tanaka,N.: "Type linterferons are essential mediators of apoptotic death in virally-infected cells." Genes to Cells. 3. 29-37 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nozawa,H.: "Functionally inactivating point mutation in the tumor suppressor IRF-1 gene was identified in human gastric cancer." Int.J.Cancer. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tanaka,T.: "Acute hepatitis caused by sexual or household transmission of GBV-C." J.Hepatology. 27. 1110-1112 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondo,S.: "Antisense telomerase treatment;induction of two distinct pathways,apoptosis and differentiation." FASEB Journal. in press. (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondo,S.: "FADD gene therapy for malignant gliomas in vitro and in vivo." Human Gene Therapy. in press. (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondo,S.: "Retrovial Transfer of Cpp32 β Gene into Malignant Gliomas in vitro and in vivo." Cancer Research. 58. 962-967 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Shimmoto,M.: "Generation and characterization of transgenic mice expressing a human mutant alpha-galactosidase with an R301Q substitution causing a variant form of Fabry disease" FEBS Letter. 417. 89-91 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ishii,S.: "α-Galactosidase transgenic mouse:Heterogeneous gene expression and posttranscriptional glycosilation in tissues." Glycoconjugate Jorunal. in press. (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 小原道法: "Modem Physician" C型肝炎とG型肝炎検査法の進歩, 5 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 小原道法: "南江堂" C型肝癌発生とウイルス増殖, 4 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Shimizu,M.Kohara,H.Yoshikra et.al.: "Detection of Interacellular Virus Particles by Electron Microscopy" Hepatology. 23. 205-209 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Kashiwakuma,A.Hasegawa,M.Kohara et.al.: "Detection of hepatitis C Virus specific core protein in serum of patients by a sensitive fluorescene enzyme immunoassay" J.Immunological Methods. 190. 79-89 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] E.Orito,M.Mizokami,M.Kohara et.al.: "Quantification of serum hepatitis C Virus cone protein level in patients chronically infected with different hepatitis C Virus genotype" GUT. 39. 876-880 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] G.Barda,M.Kohara,C.Brechot et.al.: "Hepatitis C Virus Core Protein shows a Cyfoplasmic Localization and Associates to Cellular Lipid Storage Droplets." Proc.Natl.Acad.Sci.USA. 94. 1200-1205 (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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