| Project/Area Number |
06671219
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General surgery
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| Research Institution | KYOTO PREF.UNIV.OF MEDICINE |
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Principal Investigator |
OHMORI Yoshihiro (1995) KYOTO PREF.UNIV.OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (30106405)
荒川 幸平 (1994) 京都府立医科大学, 医学部, 助手 (20167993)
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| Co-Investigator(Kenkyū-buntansha) |
大森 吉弘 京都府立医科大学, 医学部, 助教授 (30106405)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | IL-2 / AP-1 / NF-AT / POST RENAL TRANSPLANT PATIENTS / IMMUNOSUPRESSIV EEFFECTS / NF-AT / 腎移植後 / 免疫抑制剤 / IL-2遺伝子 / 免疫学的寛容 / ゲル移動度テスト法 |
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| Research Abstract |
Interleukin-2 (IL-2) produced by helper T cells has been showm to play an important role in the immunomodulation after transplantation and regulatory elements of IL-2 gene have two AP-1 and NF-AT binding sites. Recently it has been reported that binding activities of transcription factor AP-1 and NF-AT and NF-AT of anergic T cell are decreased and down-regulated as compared with normal T cells. In this study, we have assessed DNA-binding activities of AP-1 and NF-AT by electrophoretic mobility shift assay in posttransplant recipients in order to evaluate the immunomodulatory effect of the immunosuppressants.
Lymphocytes were prepared from perlpheral blood of four normal volunteers, four renal transplant recipients surviving for more than five years and four that surviving less than 5 years by gradient centrifugation over Ficoll and cultured for four hours at 37ﾟC with or without stimulation of PMA and PHA.Nuclear extracts were prepared from lympocytes by the modified method of Dignam et
… More
al. Endlabeled DNA fragments of AP-1 and NF-AT binding consensus sequences of IL-2 gene promoter regions were reacted with nuclear extracts. Electrophoretic mobility shift assays were performed and BAS-2000 Bioimage analyzer was used for the detection of DNA binding complex of AP-1 and NF-AT.Four normal volunteers showed DNA-binding activities of AP-1 and NF-AT in moderate level, but four long term surviving renal transplant recipients showed much lower level of the activities. Furthermore, a stimulatory effect of PMA and PHA was examined in these lymphocytes ; whereas all of four long term surviving recipients showed no enhancement of binding activities of AP-1 and NF-AT,two of four normal volunteers showed enhancements.
In this paper, we have shown that the DNA-binding activities of AP-1 and NF-AT in immumosuppressive state, are decreased and not enhanced even after the stimulation of PMA and PHA.Investigations of the binding activivies of AP-1 and NF-AT will provide us the estimation of immunosuppressive effect, leading to establishment of more usuful immunosuppressive regimen. Less
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