2001 Fiscal Year Final Research Report Summary
Study of the regulation of granzyme B mRNA in CTL and its clinical significance
| Project/Area Number |
11672298
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Kobe University |
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Principal Investigator |
RYO Ryukichi Kobe University, School of Medicine, Professor, 医学部, 教授 (00159237)
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| Co-Investigator(Kenkyū-buntansha) |
HORIE Osamu Kobe University, School of Medicine, Assistant Professor, 医学部, 助手 (50304118)
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| Project Period (FY) |
1999 – 2000
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| Keywords | granzyme B / perforin / apoptosis / HLA / ELISA / CTL / herpssvirus saimiri / minor histocompatibility antigen |
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| 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.
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