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Epstein-Barr virus mutagenesis by homologous recombination

Research Project

Project/Area Number 05670286
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Virology
Research InstitutionNihon University

Principal Investigator

FUJIWARA Shigeyoshi  Nihon University, School of Medicine, Lecturer, 医学部, 講師 (30173488)

Co-Investigator(Kenkyū-buntansha) ONO Yasushi  Nihon University, School of Medicine, Professor, 医学部, 教授 (30004675)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsEpstein-Barr Virus / Homologous Recombination / Recombinant Virus / Hygromycin / MT-2 / BZLF1 / Persistent Infection / EBウイルス / 変異ウイルス / 持続感染系
Research Abstract

The primary goal of this project is to generate Epstein-Barr virus (EBV) recombinants lacking the function of the EBNA3C gene and to characterize their biological activities. A targeting vector pTE3C-Hyg was constructed to insert a hygromycin resistance gene into the EBNA3C gene. This vector was transfected to the EBV-producer line B95-8 and the recombinant viruses released from the transfected B95-8 cells were inoculated to either a human EBV-negative B-cell line BJAB or cord blood mononuclear cells. Recombinant viruses were recovered in either BJAB subclones or lymphoblastoid cell lines that grew in the presence of hygromycin. The viruses recovered in these BJAB clones were shown to be the products of homologous recombination in the EBNA3C gene, whereas those recovered in lymphoblastoid lines were shown to be random recombinants. We are currently characterizing the biological activities of these recombinant viruses.
We used EBV recombinants also as a tool to generate a new experimental system of persistent EBV infection. Infection of the human T-cell line MT-2 with these EBV recombinants and subsequent selection with hygromycin permitted isolation and long-term maintenance of EBV-infected MT-2 clones. The latent EBV gene expression in these EBV-infected MT-2 clones was shown to be similar to that observed in peripheral human T-cell lymphomas. Thus these clones could be useful as a model of EBV gene regulation in T-cell lymphomas. Besides the latent genes, EBV-infected MT-2 clones constitutively express the BZLF1 gene, an immediate-early gene that triggers the productive viral cycle in B lymphocytes. Since EBV production was not detected in these MT-2 clones, we speculate that EBV gene regulation may be different between B- and T-lymphocytes.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Shigeyoshi Fujiwara: "Isolation of Epstein-Barr virus-infected clones of the human T-cell line MT-2 : use of recombinant" Journal of Virology. 69. 3900-3903 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shigeyoshi Fujiwara: "Repetitive sequence in the Epstein-Barr virus EBNA-3C gene is related to a family of minisatellite" Virus Genes. 11. 31-35 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshihiro Suzuki: "Oxidative stress triggers tyrosine phosphorylation in B cells through a redox-and inflammatory" Immunology. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasuko Nagatsuka: "Production of human monolonal antibodies to the i blood group antigen by EBV-induced" Immunology Letters. 46. 93-100 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasuko Nagatsuka: "Anti-bacterial antibodies in Epstein-Barr virus (EBV)-transformed oligoclonal B-cell lines" Letters in Applied Microbiology. 19. 206-209 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasuko Nagatsuka: "Anti-glycolipid antibodies produced by Epstein-Barr virus (EBV)-transformed oligoclonal" Human Antibodies and Hybridomas. 5. 183-186 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujiwara, S.and Ono, Y.: "Isolation of Epstein-Barr virus-infected clones of the human T-cell line MT-2 : use of recombinant viruses with a positive selection marker" J.Virol. 69. 3900-3903 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujiwara, S.and Ono, Y.: "Repetitive sequence in the Epstein-Barr virus EBNA-3C gene is related to a family of minisatellite arrays in the human genome." Virus Genes. 11. 31-35 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Suzuki, Y., Ohsugi, K., and Ono, Y.: "Oxidative stress triggers tyrosine phosphorylation in B cells through a redox- and inflammatory cytokine-sensitive mechanism" Immunology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagatsuka, Y., Watarai, S., Yasuda T., Higashi, H., Yamagata, T., and Ono, Y.: "Production of human monoclonal antibodies to the i blood group antigen by EBV-induced transformation : possible presence of a new glycolipid in cord red cell membrane and human hematopoietic cell lines" Immunology Letters. 46. 93-100 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagatsuka, Y., Hanazawa, S., Kuroiwa, Y., Fukuda, T., Suganuma, T., and Ono, Y.: "Anti-bacterial antibodies in Epstein-Barr virus (EBV) - transformed oligoclonal B-cell lines established from normal persons and autoimmune disease patients." Lett.Appl.Microbiol.19. 206-209 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagatsuka, Y., Hanazawa, S., Yasuda, T., and Ono, Y.: "Antiglycolipid antibodies produced by Epstein-Barr virus (EBV) - transformed oligoclonal B cell lines obtained from normal persons and autoimmune disease patients" Hum.Antibod.Hybridomas. 5. 183-186 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujiwara, S.: "Oncogenes." J.Nihon Univ.Med.Ass.52. 647-653 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kosaka, K., Tanaka, T., Uno, M., and Ono, Y.: "Expression of CD21 (EBV receptor) and susceptibility to EBV infection in T-cell lines." J.Nihon Univ.Med.Ass.52. 681-688 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasuko Nagatsuka: "Anti-bacterial antibodies in Epstein-Barr virus(EBV)-transformed oligoclonal B-cell lines established from normal persons and autoimmune disease patients" Letters in Applied Microbiology. 19. 206-209 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yasuko Nagatsuka: "Anti-glycolipid antibodies produced by Epsein-Barr virus(EBV)-transformed oligoclonal B-cell lines obtained from normal and autoimmune disease patients" Human Antibodies and Hybridomas. 5. 183-186 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yasuko Nagatsuka: "Production of human monoclonal antibodies to i blood group antigens by EBV-induced transformation" Immunology Letters. 44(In Press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shigeyoshi Fujiwara: "Isolation of Epstein-Barr virus-infected clones of the human T-cell line MT-2:use of recombinant viruses with a positive selection marker" Journal of Virology. 69(In Press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 小坂,和子: "T細胞株におけるEpstein-Barr Virus receptor(CD21)の発現と感染性について" 日大医学雑誌. 52. 681-688 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 藤原成悦: "がん遺伝子概観" 日大医学雑誌. 52. 647-653 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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