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

我们最近建立了一个APL细胞系(HT93)，它具有在全反式维甲酸和人类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的能力。在全反式维甲酸作用下，细胞出现核碎裂，部分出现中性粒细胞和嗜酸性粒细胞次级颗粒。在全反式维甲酸(ATRA)和粒细胞集落刺激因子(G-CSF)的作用下，HT93细胞呈中性粒细胞分化，并伴有表面CD11b的表达，NAP呈阳性反应。在ATRA和GM-CSF、IL-3或IL-5的共同作用下，HT93在第8天时表现出明显的嗜酸性粒细胞成熟，并表达嗜酸细胞过氧化物酶和主要基础蛋白。我们还观察了ATRA和二甲基亚砜(DMSO)对人髓系白血病细胞系(HL6O、HT93)粒细胞分化过程中SHP-1表达和磷酸酶活性的调节。全反式维甲酸诱导两种细胞分化与标记…相关更多的是抑制生长和S相减少。二甲基亚砜仅对HL60细胞有诱导作用，未见明显的生长抑制和S相减少。SHIP-1的表达和磷酸酶活性只有在两种细胞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-1参与了常见的髓系分化，并且SHP-1的上调并不总是与髓系细胞的生长有关。我们证明了不成熟嗜碱性粒细胞系KU812-F诱导的过氧化物酶活性的特征。免疫细胞化学显示，KU812-F细胞被抗嗜酸细胞过氧化物酶抗体染色，Northern杂交和RT-PCR检测到嗜酸细胞过氧化物酶mRNA，而不是髓过氧化物酶。嗜酸细胞过氧化物酶可能是嗜酸性粒细胞和嗜碱性粒细胞共有的分子之一。较少

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