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（HT 93），它有能力分化为中性粒细胞和嗜酸性粒细胞的反应ATRA和人类造血细胞因子。当细胞与全反式维甲酸培养时，细胞显示核分裂，部分细胞出现中性粒细胞和嗜酸性粒细胞的次级颗粒。在ATRA和G-CSF的存在下，细胞表现出多形核中性粒细胞分化，伴随表面CD 11b的表达，NAP活性阳性。在ATRA和GM-CSF、IL-3或IL-5的培养物中，HT 93在第8天表现出显著的嗜酸性粒细胞成熟，以及嗜酸性粒细胞过氧化物酶和主要碱性蛋白的表达。我们还研究了SHP-1在人髓系白血病细胞系（HL 60，HT 93）的粒细胞分化过程中对ATRA或二甲基亚砜（DMSO）的反应。ATRA诱导两种细胞系的分化，这与标记 ...更多信息 艾德生长抑制和S期减少。而DMSO仅在HL 60细胞中诱导其凋亡，对HL 60细胞生长无明显抑制作用，S期细胞数无明显减少。SHIP-1的表达和磷酸酶活性的上调，只有当两个细胞系分化为粒细胞。这些结果表明，SHP-1参与了普通髓样分化，并且SHP-1的上调并不总是与髓样细胞的生长有关。我们已经证明了在未成熟嗜碱性粒细胞系KU 812-F中诱导的过氧化物酶活性的特征。过氧化物酶活性在颗粒中以及在核周间隙和内质网中被检测到。免疫细胞化学显示，KU 812-F细胞经抗嗜酸性粒细胞过氧化物酶抗体染色，北方杂交和RT-PCR检测到嗜酸性粒细胞过氧化物酶mRNA而非髓过氧化物酶，嗜酸性粒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。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