Hiddemann W, Zühlsdorf M, Wörmann B, Koenigsmann M, Büchner T
Department of Internal Medicine, University of Münster, F.R.G.
Leuk Res. 1990;14(4):347-52. doi: 10.1016/0145-2126(90)90162-3.
In a clinical phase II trial GM-CSF was applied to 23 patients with acute leukemias carrying a high risk of early death because of old age or advanced disease. Concomitant laboratory investigations included the analysis of colony assays for normal granulopoietic progenitors (CFU-GM) and leukemic CFU-L with and without the addition of GM-CSF, as well as DNA measurements by flow cytometry (FCM) for the detection of DNA-aneuploidies. The present analysis is focused on one particular patient with acute myeloid leukemia (AML) in whom the detection of a DNA aneuploidy provided the readily accessible means to monitor the response of leukemic blasts to induction chemotherapy and subsequent GM-CSF treatment in vivo complementing the colony assay analyses performed in vitro. Prior to therapy 60% of cells revealed a DNA aneuploidy with a DNA index of 1.26 and in-vitro exposure of leukemic bone marrow blasts to GM-CSF (100 U/ml) enhanced the growth of CFU-L and CFU-GM with a significantly higher stimulatory index for CFU-L (1:19 versus 1:5.5). Three days after the completion of two cycles of induction therapy with thioguanine, cytosine arabinoside and daunorubicin (TAD 9) a morphologic bone marrow evaluation revealed aplasia without leukemic blasts. By FCM DNA analysis, however, 8% residual aneuploid cells were found. No growth of CFU-L was observed at this time point neither spontaneously nor after the addition of GM-CSF which induced the growth of CFU-GM, only. In-vivo application of GM-CSF (250 micrograms/mg2/day by continuous 24-h infusion) led to the recovery of normal granulopoiesis without evidence of a concomitant stimulation of aneuploid leukemic cells. These data indicate a change in the susceptibility of leukemic blasts to GM-CSF before and after chemotherapy. A reduction of the leukemic cell burden below a critical level or a selection of GM-CSF non-responsive early leukemic precursor cells may account for these observations.
在一项临床II期试验中，将粒细胞-巨噬细胞集落刺激因子（GM-CSF）应用于23例因年老或疾病晚期而具有早期死亡高风险的急性白血病患者。同时进行的实验室检查包括分析添加和不添加GM-CSF时正常粒细胞祖细胞（CFU-GM）和白血病CFU-L的集落测定，以及通过流式细胞术（FCM）进行DNA测量以检测DNA非整倍体。本分析聚焦于一名急性髓系白血病（AML）患者，在该患者中，DNA非整倍体的检测提供了一种易于获取的手段，用于监测白血病原始细胞对诱导化疗及后续GM-CSF体内治疗的反应，以补充体外进行的集落测定分析。治疗前，60%的细胞显示DNA非整倍体，DNA指数为1.26，白血病骨髓原始细胞体外暴露于GM-CSF（100 U/ml）可增强CFU-L和CFU-GM的生长，CFU-L的刺激指数显著更高（1:19对1:5.5）。在用硫鸟嘌呤、阿糖胞苷和柔红霉素（TAD 9）完成两个诱导治疗周期后的三天，形态学骨髓评估显示骨髓再生障碍，无白血病原始细胞。然而，通过FCM DNA分析，发现了8%的残留非整倍体细胞。此时，既未观察到CFU-L的自发生长，添加GM-CSF后也未观察到CFU-L的生长，GM-CSF仅诱导了CFU-GM的生长。GM-CSF的体内应用（通过24小时持续输注，剂量为250微克/平方米/天）导致正常粒细胞生成恢复，且无伴随非整倍体白血病细胞刺激的证据。这些数据表明化疗前后白血病原始细胞对GM-CSF的敏感性发生了变化。白血病细胞负担降低至临界水平以下或选择对GM-CSF无反应的早期白血病前体细胞可能解释了这些观察结果。
