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The effects of hematopoietic cytokines, and the changes of transcriptional factors during the proliferation, differentiation, and maturation of basophilic and eosinophilic progenitors

嗜碱性和嗜酸性祖细胞增殖、分化和成熟过程中造血细胞因子的作用和转录因子的变化

基本信息

批准号：
09671102
负责人：
FURUKAWA Tatsuo
金额：
$ 1.92万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

We recently established an APL cell line (HT93), which has a capacity to differentiate into neutrophils and eosinophils in response to ATRA and human hematopoietic cytokines. When the cells were cultured with ATRA, cells showed nuclear segmentation and developed secondary granules of neutrophils and eosinophils in part. In the presence of ATRA and G-CSF, cells showed polymorphonuclear neutrophil differentiation, accompanied by expression of surface CD11b, positive activity for NAP.In the cultures with ATRA and GM-CSF, IL-3 or IL-5, HT93 showed remarkable eosinophil maturation at day 8 as determined by luxol fast blue staining, and expression of eosinophil peroxidase and major basic protein.We also investigated the modulation of the expression and phosphatase activity of SHP-1 during granulocytic differentiation of human myeloid leukemia cell lines (HL6O, HT93) in response to ATRA or dimethylsulfoxide (DMSO). ATRA induced differentiation in both cell lines which was associated with mark … More ed growth inhibition and S-phase reduction. On the other hand, DMSO induced it only in HL60 without obvious growth inhibition and S-phase reduction. The expression and phosphatase activity of SHIP-1 were upregulated only when the two cell lines were differentiated to granulocytes. Furthermore, the changes were not dependent on the inducers or the growth inhibition, These findings suggest that SHP-1 is involved in common myeloid differentiation, and that upregulation of SHP-1 is not always related to myeloid cell growth.We have demonstrated the characteristics of peroxidase activity induced in the immatuer basophils cell line KU812-F.Ultrastructually, peroxidase activity was detected in granules as well as in the perinuclear space and endoplasmic reticulum. Immunocytochemistry revealed that KU812-F cells were stained by anti-eosinophil peroxidase antibodies, and eosinophil peroxidase mRNA, not myeloperoxidase, was detected in the cells using Northern hybridization and RT-PCR.Eosinophil peroxidase can be one of the molecules shared with eosinophils and basophils. Less

我们最近建立了一种APL细胞系（HT93），该细胞系能够在ATRA和人造血细胞因子的作用下分化为中性粒细胞和嗜酸性粒细胞。用ATRA培养细胞时，细胞出现核分裂，部分出现中性粒细胞和嗜酸性粒细胞次级颗粒。在ATRA和G-CSF存在下，细胞表现为多核中性粒细胞分化，并伴有表面CD11b的表达，NAP活性呈阳性。在ATRA和GM-CSF、IL-3或IL-5的培养中，HT93在第8天通过luxol快速蓝染色、嗜酸性过氧化物酶和主要碱性蛋白的表达，显示出明显的嗜酸性细胞成熟。我们还研究了SHP-1在人髓性白血病细胞系（hl60, HT93）粒细胞分化过程中对ATRA或二甲亚砜（DMSO）的表达和磷酸酶活性的调节。ATRA诱导两种细胞系分化，并伴有明显的生长抑制和s期减少。另一方面，DMSO仅在HL60中诱导，无明显生长抑制和s期减少。SHIP-1的表达和磷酸酶活性只有在两种细胞系分化为粒细胞时才上调。此外，这些变化不依赖于诱导剂或生长抑制。这些发现表明，SHP-1参与了常见的髓细胞分化，而SHP-1的上调并不总是与髓细胞生长有关。我们已经证明了在未成熟的嗜碱性细胞系KU812-F中诱导过氧化物酶活性的特征。超微结构上，颗粒以及核周间隙和内质网中检测到过氧化物酶活性。免疫细胞化学结果显示，KU812-F细胞被抗嗜酸性粒细胞过氧化物酶抗体染色，并通过Northern杂交和RT-PCR检测到嗜酸性粒细胞过氧化物酶mRNA，而不是髓过氧化物酶。嗜酸性粒细胞过氧化物酶是嗜酸性粒细胞和嗜碱性粒细胞共有的分子之一。少