| Project/Area Number |
11694314
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Endocrinology
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| Research Institution | Keio University |
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Principal Investigator |
SHIMIZU Nobuyoshi Keio University, School of Medicine, Professor, 医学部, 教授 (50162706)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAYANAGI Atsushi Keio University, School of Medicine, Research Associate, 医学部, 助手 (80245464)
MINOSHIMA Shinsei Keio University, School of Medicine, Associate Professor, 医学部, 助教授 (90181966)
KUDOH Jun Keio University, School of Medicine, Assistant Professor, 医学部, 専任講師 (80178003)
NAGAFUCHI Seiho Kyusyu University, School of Health Science, Professor, 医療技術短期大学, 教授 (00150441)
SAKAGUCHI Shimon Kyoto University, School of Medicine, Professor, 再生医科学研究所, 教授 (30280770)
陳 嘉竝 慶應義塾大学, 医学部, 助手 (10296725)
浅川 修一 慶應義塾大学, 医学部, 助手 (30231872)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥12,800,000 (Direct Cost: ¥11,600,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2001: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2000: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1999: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | APECED / APS-1 / AIRE / Autoimmune disease / Thymus / Antigen presenting cells / CBP / Dendritic cells / SV40T抗原 / トランスジェニックマウス / 免疫応答 / RelB / 転写調節因子 |
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| Research Abstract |
1. AIRE gene expression: We have investigated the expression of the AIRE gene by mRNA in situ hybridization and immunohistochemistry in various human tissues. We showed that AIRE is expressed in distinct cells in thymus medulla, and also in rare cells in lymph node paracortex and medulla, and in spleen and fetal liver. By RT-PCR AIRE mRNA was detected not only in thymus, lymph node, spleen and testis but also in most human and mouse tissues analyzed. We also found that the AIRE gene is restrictively expressed in the peripheral monocyte/dendritic cell lineage.
2. Structure and function of AIRE protein: We showed that AIRE fused to a heterologous DNA binding domain activates transcription from a reporter promoter. We also showed that AIRE forms homodimers through the HSR domain and interacts with common transcriptional co-activator CREB-binding protein (CBP) in vitro and in yeast nuclei through the CH1 and CH3 conserved domains of CBP.
3. AIRE gene mutations in APECED patients: We have identified four novel AIRE gene mutations in two Japanese families with APECED.
4. Establishment of AIRE-expressing cell line: To establish permanent lines of "AIRE-expressing cells", we produced a transgenic mouse expressing SV40 T-antigens driven by mouse AIRE gene promoter. We isolated cells from thymus of the transgenic mouse and maintained them in culture from which we established immortalized cells. These immortalized AIRE cells are extremely useful in studies on the molecular aspects of AIRE gene functions.
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