Two populations of erythroid cell progenitors in paroxysmal nocturnal hemoglobinuria.

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.