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Study of the regulation of granzyme B mRNA in CTL and its clinical significance

Research Project

Project/Area Number 11672298
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionKobe University

Principal Investigator

RYO Ryukichi  Kobe University, School of Medicine, Professor, 医学部, 教授 (00159237)

Co-Investigator(Kenkyū-buntansha) HORIE Osamu  Kobe University, School of Medicine, Assistant Professor, 医学部, 助手 (50304118)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
Keywordsgranzyme B / perforin / apoptosis / HLA / ELISA / CTL / herpssvirus saimiri / minor histocompatibility antigen / TNFα / Herpesvirus saimiri / granzymeB / CD8+リンパ球 / IL-12 / GST融合タンパクシステム / limiting dilution
Research Abstract

1. We reported that the patients with post-transfusion graft-versus-host disease (PT-GVHD) had a partial remission after administration of nafmostat mesilate, a serine protease inhibitor, thus indicating that granzyme(gr) plays an important role in organ failure in PT-GVHD.
2. Cytokine dependency of granzyme B(grB) mRNA expression was investigated using competitive RT-PCR. We found that the combination of IL-2 and IL-12 additively increased the expression of grB mRNA. It therefore seems possible that YT-u may have a different intracellular signal transudation system for cytokine-induced regulation of grB mRNA from that of normal NK cells.
3. Six kinds of monoclonal antibodies which recognized three different epitope were prepared using the recombinant grB protein produced with the GST fusion protein system. One step ELISA for grB was established using these antibodies. We clarified that this ELISA was two thousands as sensitive as an enzyme assay.
4. CTL clone was established in HLA- homozygous pairs and HLA-heterozygous pairs in vitro. To obtain a stable CTL clone, immortalization of Tcell was performed by infection with Herpesvirus saimiri. We found little difference between TNF production and the induction of grB gene expression in two CTLs which recognized major and minor histocompatibility antigen. We elucidated the importance of minor histocompatibility antigen in the induction of CTL clone.
5. To obtain an active grB and perforin, the experiment of the protein expression system with insect cells is now in progress.

Report

(3 results)
  • 2001 Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Ryukichi Ryo, et al.: "Treatment of post-transfusion graft-versus-host disease with nafmostat mesilate, a serine protease inhibitor"Vox Sanguinis. 76. 241-246 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katsuyasu Saigo, Ryukichi Ryo: "Therapeutic Strategy for post-transfusion graft-vs-host disease"International Journal of Hematology. 69. 147-151 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Osamu Horie, Ryukichi Ryo: "Establishment of a lymphocyte clone expressing granzyme B and perforin mRNAs immortalized with Herpesvirus saimiri"Bulletin of Health Sciences Kobe. 16. 39-44 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Osamu Horie, Ryukichi Ryo: "Cytekine dependency of granzyme B mRNA expression in natural killer leukemic cell, YT-U"Bulletin of Health Sciences Kobe. 16. 57-62 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ryo R., Saigo K., Hashimoto M, Kohsaki M, et al: "Treatment of post-transfusion grant-versus-host disease with nafmostat mesilate, a serine protease inhibitor"Vox Sanguinis. 76-4. 241-246 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saigo K., Ryo R.: "Therapeutic strategy for post-transfuison graft-vs.-host disease"International Journal of Hematology. 69-3. 147-151 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Horie O., Ryo R.: "Establishment of a lymphocyte clone expressing granzyme B and perforin mRNAs immortalized with Herpesvirus saimiri"Bulletin of Health Sciences Kobe. 16. 39-44 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Horie O., Ryo R.: "Cytokine dependency of granzyme B mRNA expression in natural killer leukemic cell line, YT-u"Bulletin of Health Sciences Kobe. 16. 57-62 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ryukichi Ryo, et al.: "Treatment of post-transfusion grafe-versus-host disease with nafmostat mesilate, a serine protease inhibitor"Vox Sanguinis. 76・4. 241-246 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Katsuyasu Saigo, et al.: "Therapeutic Strategy for post-transfusion grafe-vs.-host disease"International Journal of Hematology. 69・3. 147-151 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Osamu Horie et al.: "Establishment of a lymphocyte clone expressing granzyme B and perforin mRNAS immortalized with Herpesvirus saimiri"Bulletin of Health Science Kobe. 16. 39-44 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Osamu Horie et al.: "Cytokine dependency of granzyme B mRNA expression in natural killer leukemic cell line, YT-u."Bulletin of Health Science Kobe. 16. 57-62 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 富和洋子、寮隆吉: "Herpesvirus saimiri を用いた細胞障害性Tリンパ球クロ-ン樹立に関する研究"神戸大学医学部保健学科検査技術科学専攻卒業論文集. 1. 77-80 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 濱田敬子、寮隆吉: "各種血球のGranzyme B mRNA発現とそれに対するcytokineの影響"神戸大学医学部保健学科検査技術科学専攻卒業研究論文集. 1. 81-84 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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