Proliferation and differentiation of granulocytes

造血细胞的增殖、分化和激活受一种称为集落刺激因子(CSF)的糖蛋白家族的调节。粒细胞集落刺激因子(G-CSF)是脑脊液(CSF)的一员，能刺激中性粒细胞的增殖和分化。在祖细胞分化为中性粒细胞的过程中，髓过氧化物酶等几种酶被特异性地诱导。我们以前已经分离了人和小鼠G-CSF的cDNA，并阐明了G-CSF染色体基因的结构组织。此外，还从人早幼粒白血病HL-60细胞中分离到人髓过氧化物酶基因。在本项目中，我们利用重组DNA技术在小鼠C127细胞中产生了人和小鼠的G-CSF。纯化的重组人粒细胞集落刺激因子能刺激小鼠髓系白血病细胞增殖，并能诱导WEHI-3BD^+细胞向单核细胞和粒细胞分化。当Pur…进一步皮下注射重组G-CSF，小鼠可见明显的粒细胞生成和脾肿大。用~(125)I标记的G0CSF对G-CSF的细胞受体进行了分析，结果表明，几个髓系白血病细胞的G-CSF受体的解离常数为100-200 pM，每个细胞有1000-2000个拷贝此外，该受体与[125I]G-CSF的交联表明，该受体的相对分子质量为140,000-150,000。利用髓过氧化物酶的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。小鼠髓系白血病细胞NFS-60和WEHI-3BD^+细胞表达髓过氧化物酶，而G-CSF只下调WEHI-3BD^+细胞髓过氧化物酶的表达，而不下调其表达。G-CSF能诱导WEHI-3BD^+细胞分化，但不能诱导NFS60细胞分化，提示髓过氧化物酶基因可作为中性粒细胞分化的标志物。分离了人类染色体髓过氧化物酶基因，并证明其由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，其中一个似乎编码嗜酸性粒细胞过氧化物酶。较少

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