| Project/Area Number |
05670157
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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 |
Pathological medical chemistry
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| Research Institution | Osaka Bioscience Institute |
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Principal Investigator |
SUDA Takashi 1st Department, Osaka Bioscience Institute Scientist, 第1研究部, 研究員 (70250090)
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| Co-Investigator(Kenkyū-buntansha) |
NAGATA Shigekazu 1st Department, Head, 第1研究部, 部長 (70114428)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Fas ligand / Fas / apoptosis / autoimmune disease / CTL / gld mutation / cDNA cloning / chromosomal mapping / T細胞 / Fas抗原 / 細胞傷害性T細胞 / 精製 |
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| Research Abstract |
Fas is a 45 kD cell-surface protein belonging to the TNF receptor family. Some mAbs recognizing Fas (anti-Fas and anti-Apol) induce apoptotic cell death in cells expressing Fas. These facts suggested that Fas is a receptor for a death factor. We succeeded to purify the natural ligand for Fas(FasL) from the membrane fraction of sublines of CTL hybridoma (rat x mouse) PC60-d10S by Fas-Fc (a fusion protein consisting of the extracellular portion of Fas and the Fc portion of human IgG1) affinity column. The purified FasL was a membrane glycoprotein having a Mr of 40 kD.Subsequently, a cDNA encoding FasL was a membrane glycoprotein having a Mr of 40 kD.Subsequently, a cDNA encoding FasL was isolated from a cDNA library derived from the same cell line by panning method using Fas-Fc. The deduced amino acid sequence of its cDNA indicated that FasL is a type II transmembrane protein belonging to the TNF family. Since the isolated FasL cDNA was rat origin, we isolated its human and mouse homolog
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ues using rat FasL cDNA as a probe. The amino acid identities of FasLs between human-mouse, human-rat and mouse-rat are 76.9%, 75.8% and 91.4%, respectively. Human and mouse FasL are equally potent to induce apoptosis in cells expressing human or mouse Fas, indicating that the Fas-FasL interaction is fully cross-reactive between human and mouse. Expression of FasL mRNA was restricted to certain tissues including lymphoid organs (thymus, spleen, lymph nodes), lung and small intestine, and to cell lines of the T cell lineage. These results suggest that FasL plays important roles in both systemic and mucosal immunity. We have previously demonstrated that mouse lpr mutation is a loss of function mutant of Fas gene. In this research, we showed that mouse gld mutation is a loss of function mutant of FasL gene. Since these mice develop lymphadenopathy by the accumulation of abnormal CD4-CD8-T cells and suffer from SLE-like autoimmune diseases, it is likely that the Fas/FasL system is involved in the apoptosis of T cells upon turnover of activated T cells and/or clonal deletion of autoreactive T cells. Less
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