| Project/Area Number |
12670298
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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 |
Immunology
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| Research Institution | Nagoya University |
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Principal Investigator |
SUZUKI Haruhiko Graduate School of Medicine, Nagoya University, Associate Professor, 大学院・医学研究科, 助教授 (90283431)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASHIMA Izumi Graduate School of Medicine, Nagoya University, Professor, 大学院・医学研究科, 教授 (40022826)
加藤 昌志 名古屋大学, 医学部, 助教授 (10281073)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Resting T cells / Lymph node-derived stromal cells / Survival factor / Cytokine / Gene cloning / Functional assay / Gene-targeted mice / Chemokine / T細胞生存因子 / 遺伝子ターゲティング / リンパ節ストロマ細胞 / cDNAクローニング |
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| Research Abstract |
(1) Establishment of in vitro culture system to support the survival of mature resting T cells and analysis of its mechanism in effective action.
We succeeded in establishing the culture of stromal cells derived from lymph nodes of IL-2 receptor β-knock out mice. Using these cells, we analyzed the mechanism how mature resting T cells survive. Lymph nodes-derived stromal cells effectively supported the survival of t cells, and it was suggested that the survival was mediated by some liquid-soluble factors. IL-7 and other cytokines also supported T cell survival but induced proliferation at the same time. In contrast, the most important feature of lymph node-derived stromal cells was that they did not induce proliferation at all, but supported the T cell survival with keeping its resting state.
(2) Gene cloning of factors supporting the survival of resting T cells.
We cloned the genes of factors that supported T cell survival by screening cDNA from lymph node-derived stromal cells. As a result of screening approximately 100,000 clones with transfection to COS-7 cells and functional assay, 8 cDNA clones were confirmed to have a functional activity to support the T cell survival. Gene sequences of all the 8 clones had alre4ady been registered in the database, but some of them were still functionally unknown.
(3) Generation of gene-targeted mice for C-C chemokine 6 and C-C chemokine 9.
We started to generate the gene-targeted mice for C-C chemokine 6, which had been cloned in our study, and C-C chemokine 9, the gene of which was located close to C-C chemokine 6, in order to search for their functions in a whole body. The plasmids for the gene targeting of each gene were constructed and are introduced into ES cells.
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