Deschamps J F, Bodevin E, Savariau E, Enouf J, Lompre A M, Dosquet C, Caen J
INSERM U-150, Hôpital Lariboisière, Paris, France.
Exp Hematol. 1991 Sep;19(8):729-33.
We report the presence of a Ca(2+)-ATPase in human megakaryocytes (MK) using an immunofluorescence technique on bone marrow smears and especially on normal MK progenitors in culture. This finding is based on the comparative staining of MK with 1) a well-characterized antibody raised against purified rabbit skeletal sarcoplasmic reticulum Ca(2+)-ATPase, 2) antibody P2 raised against the glycoprotein IIb-IIIa complex as a marker of megakaryocytic lineage, and 3) anti-glycophorin A as a marker of erythroid lineage. On bone marrow smears, all cells recognized by P2 were also labeled with the anti-Ca(2+)-ATPase antibody. In culture, a maximum number of MK colonies was observed at day 11. From days 2-4, some MK precursors appeared stained both with the anti-Ca(2+)-ATPase and P2 antibodies; other cells were reactive with both anti-Ca(2+)-ATPase and anti-glycophorin A antibodies. From day 5 of culture, cells were either simultaneously stained with P2 and anti-Ca(2+)-ATPase antibodies or with anti-glycophorin A antibody, but not with the anti-Ca(2+)-ATPase antibody. Besides this first evidence of an early expression of a Ca(2+)-ATPase in MK, this work provides a useful tool for identification of MK by immunofluorescence.
我们运用免疫荧光技术，在骨髓涂片尤其是培养的正常巨核细胞（MK）祖细胞上，报告了人类巨核细胞中存在钙（Ca2+）-ATP酶。这一发现基于以下比较性染色：1）用针对纯化的兔骨骼肌肌浆网钙（Ca2+）-ATP酶产生的一种特征明确的抗体对巨核细胞进行染色；2）用针对糖蛋白IIb-IIIa复合物产生的抗体P2作为巨核细胞系的标志物进行染色；3）用抗血型糖蛋白A作为红系标志物进行染色。在骨髓涂片上，所有被P2识别的细胞也被抗钙（Ca2+）-ATP酶抗体标记。在培养过程中，第11天观察到最大数量的巨核细胞集落。在培养的第2至4天，一些巨核细胞前体同时被抗钙（Ca2+）-ATP酶抗体和P2抗体染色；其他细胞则与抗钙（Ca2+）-ATP酶抗体和抗血型糖蛋白A抗体均有反应。从培养第5天起，细胞要么同时被P2抗体和抗钙（Ca2+）-ATP酶抗体染色，要么被抗血型糖蛋白A抗体染色，但不再被抗钙（Ca2+）-ATP酶抗体染色。除了首次证明钙（Ca2+）-ATP酶在巨核细胞中的早期表达外，这项工作还为通过免疫荧光鉴定巨核细胞提供了一种有用的工具。
