Moore J G, Frank M M, Müller-Eberhard H J, Young N S
J Exp Med. 1985 Oct 1;162(4):1182-92. doi: 10.1084/jem.162.4.1182.
A glycoprotein that regulates the deposition of C3b on the erythrocyte surface, called decay-accelerating factor or DAF, is absent from the red blood cells (RBC) of patients with paroxysmal nocturnal hemoglobinuria (PNH), explaining in part their abnormal sensitivity to complement. We used a specific antiserum to DAF, flow microfluorometry, and clonogenic assays for erythroid progenitor cells to study PNH erythropoiesis in vitro. By fluorescence-activated cell sorter analysis, all RBC from normal individuals are DAF+. In contrast, the RBC of six patients with PNH showed discrete populations of DAF- cells (10-44%; x +/- SEM = 31 +/- 6%). The DAF- RBC population was partly eliminated by prior acidified serum lysis. To determine whether erythropoietic progenitors expressed DAF, bone marrow cells were sorted by flow microfluorometry and the separated DAF+ and DAF- populations then cultured in vitro. In two normal individuals, but also in six patients with PNH, erythroid colonies formed only from cells in the DAF+ fraction. However, a variable proportion of the normoblast progeny of these DAF+ progenitor cells from patients with PNH was DAF-. Individual bursts removed from cultures of PNH bone marrow showed two discrete populations by fluorescence; the majority of normoblasts were DAF-, only 3 of 27 individual bursts had greater than 50% DAF+ cells, and in three patients, DAF- normoblasts averaged 79%. In contrast, the progeny of individual bursts from normal individuals comprised a unimodal DAF+ population. In each PNH patient, one normal burst (greater than 80% DAF+ normoblasts) was detected, possibly reflecting a normal residual population of erythroid progenitors. By the criterion of DAF expression, there was no evidence of separate populations of normal and PNH type progenitor cells. The phenotypically normal erythroid progenitors of PNH bone marrow acquire the PNH characteristics during differentiation in vitro.
一种调节C3b在红细胞表面沉积的糖蛋白，称为衰变加速因子或DAF，在阵发性夜间血红蛋白尿（PNH）患者的红细胞（RBC）中缺失，这部分解释了他们对补体的异常敏感性。我们使用针对DAF的特异性抗血清、流式微量荧光测定法以及红系祖细胞的克隆形成试验来体外研究PNH的红细胞生成。通过荧光激活细胞分选仪分析，正常个体的所有红细胞均为DAF阳性。相比之下，6例PNH患者的红细胞显示出离散的DAF阴性细胞群体（10% - 44%；x ± SEM = 31 ± 6%）。DAF阴性红细胞群体可通过预先的酸化血清裂解部分消除。为了确定红系祖细胞是否表达DAF，通过流式微量荧光测定法对骨髓细胞进行分选，然后将分离出的DAF阳性和DAF阴性群体进行体外培养。在2名正常个体以及6例PNH患者中，红系集落仅由DAF阳性部分的细胞形成。然而，这些来自PNH患者的DAF阳性祖细胞的晚幼红细胞后代中，有可变比例为DAF阴性。从PNH骨髓培养物中取出的单个集落通过荧光显示出两个离散群体；大多数晚幼红细胞为DAF阴性，27个单个集落中只有3个的DAF阳性细胞大于50%，并且在3例患者中，DAF阴性晚幼红细胞平均占79%。相比之下，正常个体单个集落的后代构成单峰DAF阳性群体。在每例PNH患者中，都检测到一个正常集落（大于80%的DAF阳性晚幼红细胞），这可能反映了红系祖细胞的正常残余群体。根据DAF表达的标准，没有证据表明存在正常和PNH型祖细胞的单独群体。PNH骨髓中表型正常的红系祖细胞在体外分化过程中获得了PNH特征。
