Proliferation and differentiation of granulocytes

粒细胞的增殖和分化

• 批准号: 62480131
• 负责人: NAGATA Shigekazu
• 金额: $ 3.65万
• 依托单位: Osaka Bioscience Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62480131/
• 关键词: 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

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 … More 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

造血细胞的增殖、分化和活化受称为集落刺激因子（CSF）的糖蛋白家族调节。粒细胞集落刺激因子（G-CSF）是CSF的一员，刺激嗜中性粒细胞的增殖和分化。在祖细胞分化成中性粒细胞的过程中，特异性地诱导几种酶，如髓过氧化物酶。我们先前已经分离出人和小鼠G-CSF的cDNA，并阐明了G-CSF染色体基因的结构组织。此外，从人早幼粒细胞白血病HL-60细胞中分离了人髓过氧化物酶的cDNA。在这个项目中，我们使用重组DNA技术在小鼠C127细胞中生产了人和小鼠G-CSF。纯化的重组人G-CSF能刺激小鼠髓性白血病NFS-60细胞增殖，并能诱导WEHI-3BD^+细胞向单核细胞和粒细胞分化。当pur ...更多信息 将纯化的重组G-CSF皮下注射给小鼠，可观察到小鼠明显的粒系造血和脾肿大。已经使用[125 I]标记的G 0 CSF分析了G-CSF的细胞受体，并且显示在解离常数为100 - 200 pM时，几种骨髓性白血病细胞具有每个细胞1，000 - 2，000个拷贝的受体。用[125 I] G-CSF与该受体交联，表明该受体的分子量为140，000 - 150，000。利用髓过氧化物酶cDNA研究了髓过氧化物酶基因在中性粒细胞分化过程中的表达调控。小鼠髓性白血病NFS-60和WEHI-3BD^+细胞表达髓过氧化物酶mRNA，G-CSF仅下调WEHI-3BD^+细胞的髓过氧化物酶表达，但对NFS-60细胞无影响。G-CSF对WEHI-3BD^+细胞有诱导分化作用，但对NFS-60细胞无诱导分化作用，提示髓过氧化物酶基因可作为中性粒细胞分化的标志。分离出髓过氧化物酶的人类染色体基因，并显示它由12个外显子组成。在人类基因组中还存在其他髓过氧化物酶相关基因，其中一个可能编码嗜酸性粒细胞过氧化物酶。少

项目成果

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Hara,K.: Exp.Hematol.16. 256-261 (1988)

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Tani,K.: Proc.Natl.Acad.Sci.USA. 85. 1792-1795 (1988)

Tani,K.：Proc.Natl.Acad.Sci.USA。

Tsuchiya, M.: "Characterization of recombinant human granulocyte-colony stimulating factor produced in mouse cells" The EMBO J.6. 611-616 (1987)

Tsuchiya, M.：“小鼠细胞中产生的重组人粒细胞集落刺激因子的表征” EMBO J.6。

Buchberg,A.M.: Oncogene Res.2. 149-166 (1988)

Buchberg，A.M.：癌基因研究 2。