| Project/Area Number |
62480131
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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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 |
NAGATA Shigekazu Head, Department of Molecular Biology, Osaka Bioscience Institute, 第1研究部, 研究部長 (70114428)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUNAGA Rikiro Researcher, Department of Molecular Biology, Osaka Bioscience Institute, 第1研究部, 研究員 (40189965)
SAKAMAKI Kazuhiro Researcher, Department of Molecular Biology, Osaka Bioscience Institute, 第1研究部, 特別研究員 (20271017)
KAZIRO Yoshito Professor, Institute of Medical Science, University of Tokyo, 医科学研究所・, 教授 (90012690)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥4,200,000 (Direct Cost: ¥4,200,000)
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| Keywords | Haemopoiesis / Granulocyte colony-stimulating factor / Myeloid leukemia / Proliferaton and differentiaton / Neutrophil / 血球の増殖と分化 / 受容体 / ミエルペルオキシダーゼ / 好酸球 / 多能性幹細胞 / 遺伝子工学 / 細胞の増殖と分化 |
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| Research Abstract |
Proliferation, differentiation and activation of haemopoietic cells are regulated by a family of glycoprotein called colony stimulating factors (CSF). Granulocyte colony-stimulating factor (G-CSF) is a member of CSF, and stimulates proliferation and differentiation of neutrophilic granulocytes. During differentiation of the progenitor cells into neutrophils, several enzymes such as myeloperoxidase are specifically induced. We have previously isolated the cDNA for human and mouse G-CSF, and elucidated the structural organization of the G-CSF chromosomal gene. Furthermore, the cDNA for human myeloperoxidase was isolated from human promyelocytic leukemia HL-60 cells. In this project, we have produced human and mouse G-CSF in mouse C127 cells using recombinant DNA technology. The purified recombinant human G-CSF could stimulate the proliferation of mouse myeloid leukemia NFS-60 cells while it could induce the differentiation of WEHI-3BD^+ cells into monocytes and granulocytes. When the pur
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ified recombinant G-CSF was subcutaneously injected into mouse, the remarkable granulopoiesis and splenomegaly were observed in mice. The cellular receptor for G-CSF has been analyzed using [125I] labeled G0CSF, and it was shown that several myeloid leukemia cells have the receptor in 1,000 - 2,000 copies per cells at the dissociation constant of 100 - 200 pM. Furthermore, the cross-linking of the receptor with [125I] G-CSF indicated the molecular weight of the receptor is 140,000 - 150,000.
Using the cDNA for myeloperoxidase, the regulation of the expression of myeloperoxidase gene during differentiation into neutrophils was studied. Mouse myeloid leukemia NFS-60 and WEHI-3BD^+ cells express mRNA for myeloperoxidase, and the expression of myeloperoxidase was down-regulated by G-CSF in only WEHI-3BD^+ cells but not NFS-60 cells. Since G-CSF can induce differentiation of WEHI-3BD^+ cells but not NFS-60 cells, it may suggest that myeloperoxidase gene can be used as a marker for differentiation of neutrophil. The human chromosomal gene for myeloperoxidase was isolated, and shown it is consisting of 12 exons. There exist other myeloperoxidase related genes in human genome, one of which seems to code for eosinophil peroxidase. Less
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