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Gene expression of TT virus and development of its antigen and antibody assay systems

Research Project

Project/Area Number 13670303
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionJichi Medical School

Principal Investigator

NISHIZAWA Tsutomu  Jichi Medical School, Department of Infection and Immunity, Assistant Professor, 医学部, 講師 (30306112)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsTT virus / gene expression / antigen and antibody assay system / Baculovirus / Western blotting / 抗原、抗体 / 遺伝子型 / 抗体
Research Abstract

In 1997, in my laboratory, a novel unenveloped virus with a small, covalently closed circular genome of single-stranded DNA of 3.9 kilobases was discovered from the serum of a patient with transfusion-related acute hepatitis of unknown etiology, and named as TT virus (TTV) after the initial of the index patient. The aims of the present study were to obtain recombinant TTV proteins by gene expression and to develop assay systems of TTV-related antigens and antibodies. In the first fiscal year, parts of TTV ORF1 and ORF2 proteins were expressed in E.coli, and purified. Using the expressed proteins as immunogens, two distinct murine monoclonal antibodies (mAbs) were raised against ORF1 protein (anti-ORF1 mAb) and ORF2 protein (anti-ORF2 mAb). In the second fiscal year, antigenic epitopes to which anti-ORF1 mAb and anti-ORF2 mAb are directed, were determined using multipin peptides. These two mAbs were successfully applicable to Western blotting for the detection of TTV proteins. Then, for the purpose of large-scale preparation of TTV proteins, four ORFs (ORF1-ORF4) of TTV were expressed separately or in combinations in silkworms using recombinant baculovirus vectors, and tandem dimers of TTV genome were introduced into cultured hepatoma cell line (HuH7). Unexpectedly, TTV proteins were not yielded in both expression experiments, enough for the establishment of TTV antibody detection system. However, in this study, I could first develop two mAbs against TTV ORF1 and ORF2, both of which are useful for specifically detecting TTV-related antigens.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Peng YH: "Analysis of the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants"Arch Virol. 147. 21-41 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamoto H: "Genomic characterization of TT viruses (TTVs) in pigs, cats and dogs and their relatedness with species-specific TTVs in primates and tupaias"J Gen Virol. 83(6). 1291-1297 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ohto H: "TT virus infection during childhood"Transfusion. 42(7). 892-898 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asabe S: "Phosphorylation of serine-rich protein encdoded by open reading frame 3 of the TT virus genome"Biochem Biophys Res Commun. 286(2). 298-304 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamoto H: "Genomic and evolutionary characterization of TT virus (TTV) in tupaias and comparison with species-specific TTVs in humans and non-human primates"J Gen Virol. 82(9). 2041-2050 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Muljono DH: "Molecular epidemiology of TT virus (TTV) and characterization of two novel TTV genotypes in Indonesia"Arch Virol. 146. 1249-1266 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shibayama T: "Inverse relationship between the titre of TT virus DNA and the CD4 cell count in patients infected with HIV"AIDS. 15. 563-570 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tsuda F: "IgM-class antibodies to TT virus (TTV) in patients with acute TTV infection"Hepatol Res. 19. 1-11 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Peng YH et al: "Analysis of the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants"Arch Virol. 147. 21-41 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamoto H et al: "Genomic characterization of TT viruses (TTVs) in pigs, cats and dogs and their ralatedness with species-specific TTVs in primates and tupaias"J Gen Virol. 83(6). 1291-1297 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ohto H et al: "TT virus infection during childhood"Transfusion. 42(7). 892-898 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asabe S et al: "Phosphorylation of serine-rich protein encdoded by open reading frame 3 of the TT virus genome"Biochem Biophys Res Commun. 286(2). 298-304 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamoto H et al: "Genomic and evolutionary characterization of TT virus (TTV) in tupaias and comparison with species-specific TTVs in humans and non-human primates"J Gen Virol. 82(9). 2041-2050 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Muljono DH et al: "Molecular epidemiology of TT virus (TTV) and characterization of two novel TTV genotypes in Indonesia"Arch Virol. 147. 21-41 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shibayama T et al: "Inverse relationship between the titre of TT virus DNA and the CD4 cell count in patients infected with HIV"AIDS. 15. 563-570 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tsuda F et al: "IgM-class antibodies to TT virus (TTV) in patients with acute TTV infection"Hepatol Res. 19. 1-11 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Peng YH: "Analysis ot the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants"Arch Virol. 147. 21-41 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okamoto H: "Genomic characterization of TT viruses(TTVs) in pigs, cats and dogs and their relatedness with species-specific TTVs in primates and tupaias"J Gen Vilo. 83(6). 1291-1297 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ohto H: "TT virus infection during childhood"Transfusion. 42(7). 892-898 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Peng, H, Y: "Analysis of the entire genomes of thirteen TT virus variants classifiable into the fourth and fifth genetic groups, isolated from viremic infants"Arch. Virol.. 147. 21-41 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Okamoto, H: "Heterogeneous distribution of TT virus of distinct genotypes in multiple tissues from infected humans"Virology. 288. 358-368 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Asabe, S: "Phosphorylation of serine-rich protein encoded by open reading frame 3 of the TT virus genome"Biochem. Biophys. Res. Commum.. 286. 298-304 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Muljono, D, H: "Molecular epidemiology of TT virus (TTV) and characterization of two novel TTV genotypes in Indonesia"Arch. Virol.. 146. 1249-1266 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Okamoto, H: "Genomic and evolutionary characterization of TT virus (TTV) in tupaias and comparison with species-specific TTVs in humans and non-human primates"J. Gen. Virol.. 82. 2041-2050 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tsuda, F: "IgM-class antibodies to TT virus (TTV) in patients with acute TTV infection"Hapatology Res.. 19. 1-11 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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