Rotoli B, Robledo R, Scarpato N, Luzzatto L
Blood. 1984 Oct;64(4):847-51.
We have grown erythroid cell colonies from two patients with paroxysmal nocturnal hemoglobinuria (PNH). At 11 to 13 days, individual bursts were picked and incubated for 24 hours with 3H-leucine in order to label total cell protein (mainly hemoglobin). After appropriate washing, each burst was subjected to a miniaturized acidified serum test, and lysis was measured by the release of radioactivity. In bursts from normal controls, lysis was 19% +/- 13% SD. By contrast, of 58 bursts from PNH patients, 14 had lysis similar to that of controls (mean 15.4% +/- 10.6%), while 44 had lysis ranging from 42.2% to 85.8% (mean 70.3% +/- 10.4%). Colonies sensitive to acidified serum were acetylcholinesterase (AchE) negative, whereas normal colonies were AchE-positive. Thus, based on two independent criteria, a dual population of erythroid burst-forming units (BFU-E) can be demonstrated in PNH. These data confirm directly the somatic mutation model of the pathogenesis of PNH, and by these methods the relative sizes of the normal and the PNH cell populations can be measured at the level of the erythroid cell precursors.
我们从两名阵发性睡眠性血红蛋白尿（PNH）患者中培养出了红系细胞集落。在第11至13天，挑选出单个集落爆发，并与³H-亮氨酸一起孵育24小时，以便标记总细胞蛋白（主要是血红蛋白）。经过适当洗涤后，每个集落爆发都进行了小型化酸化血清试验，并通过放射性释放来测量裂解情况。在正常对照的集落爆发中，裂解率为19%±13%标准差。相比之下，在PNH患者的58个集落爆发中，14个的裂解情况与对照相似（平均15.4%±10.6%），而44个的裂解率在42.2%至85.8%之间（平均70.3%±10.4%）。对酸化血清敏感的集落乙酰胆碱酯酶（AchE）阴性，而正常集落AchE阳性。因此，基于两个独立标准，可以在PNH中证明存在双种群的红系爆式集落形成单位（BFU-E）。这些数据直接证实了PNH发病机制的体细胞突变模型，并且通过这些方法可以在红系细胞前体水平测量正常和PNH细胞群体的相对大小。
