Oskéritzian C, Le Mao J, David B
Institut Pasteur, Unite d'Immuno-Allergie, Paris, France.
Exp Hematol. 1998 Jun;26(6):472-7.
We previously reported an in vitro bioassay permissive to eosinophil production from naive mouse bone marrow cells. Based on cytosmear analysis, recombinant murine (rm) interleukin (IL)-3 combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) provided better quantitative cellular yields compared with rmIL-5. In the present study, we compared the eosinophil peroxidase (EPO) content obtained in both in vitro conditions using microtitration. Parallel with the EPO assay, the percentage of eosinophils present in cultures was determined by cytosmear analysis. Cells incubated with IL-5 displayed significantly higher EPO activity than those cultured in the presence of IL-3 and GM-CSF. At the end of the culture period, cells were further enriched in eosinophils as determined by centrifugation gradient. Enhanced amounts of cellular EPO were detected when cultures were performed in the presence of IL-5. With regard to the correlation between eosinophil maturation and EPO acquisition, eosinophil cells cultured with rmIL-5 expressed significantly more EPO activity than those generated in the presence of rmIL-3 and GM-CSF.
我们之前报道了一种允许从幼稚小鼠骨髓细胞产生嗜酸性粒细胞的体外生物测定法。基于细胞涂片分析,与重组小鼠(rm)白细胞介素(IL)-5相比,重组小鼠IL-3与粒细胞-巨噬细胞集落刺激因子(GM-CSF)联合使用可提供更好的定量细胞产量。在本研究中,我们使用微量滴定法比较了两种体外条件下获得的嗜酸性粒细胞过氧化物酶(EPO)含量。与EPO测定平行,通过细胞涂片分析确定培养物中嗜酸性粒细胞的百分比。与在IL-3和GM-CSF存在下培养的细胞相比,用IL-5孵育的细胞表现出显著更高的EPO活性。在培养期结束时,通过离心梯度测定,细胞中嗜酸性粒细胞进一步富集。当在IL-5存在下进行培养时,检测到细胞EPO量增加。关于嗜酸性粒细胞成熟与EPO获得之间的相关性,用rmIL-5培养的嗜酸性粒细胞比在rmIL-3和GM-CSF存在下产生的细胞表达显著更多的EPO活性。