Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines
Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines
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DOI：
10.1182/blood.v90.7.2555.2555_2555_2564
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发表时间：
1997-10-01
期刊：
影响因子：
20.3
通讯作者：
Hogge, DE
中科院分区：
文献类型：
--
作者：
Ailles, LE;Gerhard, B;Hogge, DE
使用H-3-胸苷（H-3-Tdr）掺入试验或甲基纤维素培养物（集落试验）中的短期增殖终点，分析白细胞介素-3（IL-3）、Steel因子（SF）和flt-3配体（FL）对急性髓性白血病（AML）母细胞的促有丝分裂活性，结果显示，超过90%的样本含有对这些细胞因子中的一种或多种有反应的细胞。利用这些信息，在有或没有这三种生长因子中的一种或多种的补充的情况下，测试了已知支持正常长期培养起始细胞（LTC-IC）的培养条件，以确定其支持来自AML患者的10个细胞样品的原始祖细胞的能力。在所有情况下，当AML外周血或骨髓细胞在预先建立的正常人骨髓饲养细胞（HMF）和/或SI/SI小鼠成纤维细胞饲养细胞上共培养5周后，检测到细胞遗传学异常集落形成细胞（CFC），并且在这些5周龄LTC中检测到的CFC数量与输入AML细胞的数量保持线性关系。对10个样本中的6个进行有限稀释分析，结果显示AML细胞启动LTC（AML LTC-IC）的频率比相同细胞样本中AML-CFC的频率低5- 300倍，而每个LTC-IC产生的CFC平均数量从1到13不等。令人惊讶的是，在每种情况下，在AML患者血液中检测到的细胞遗传学正常ITC-IC的浓度比先前在正常个体血液中观察到的浓度高至少10倍。“混合”小鼠成纤维细胞饲养细胞经工程改造，以产生人类G-CSF，IL-3和SF，并没有提高AML LTC-IC的检测，但确实增加了3/4例患者样本的LTC中细胞遗传学正常CFC的输出。用IL-3和外源性SF和/或FL补充AML LTC，9个患者样品中有5个的5周龄LTC的AML-CFC输出增加大于或等于两倍，而在一个情况下，外源性FL的添加减少了LTC的恶性CFC输出。这些研究表明，支持正常LTC-IC的条件也允许检测功能类似但罕见的AML祖细胞类型。此外，正常细胞和白血病细胞对正常LTC-IC上的各种细胞因子的反应存在差异。对这些差异的进一步分析可能会增强我们对白血病发生的理解，并导致可以用于治疗的观察结果。(C)1997年，美国血液学会。
Analysis of the mitogenic activity of interleukin-3 (IL-3), Steel factor (SF), and flt-3 ligand (FL) on acute myelogenous leukemia (AML) blasts using the short-term endpoints of proliferation in H-3-thymidine (H-3-Tdr) incorporation assays or methylcellulose cultures (colony assays) showed that greater than 90% of samples contained cells that were responsive to one or more of these cytokines. With this information, culture conditions that were known to support normal long-term culture-initiating cells (LTC-IC) were tested, with or without supplements of one or more of these three growth factors, for their ability to support primitive progenitors from 10 cell samples from patients with AML. In all cases cytogenetically abnormal colony farming cells (CFC) were detected after 5 weeks when AML peripheral blood or marrow cells were cocultured on preestablished, normal human marrow feeders (HMF) and/or SI/SI mouse fibroblast feeders and the number of CFC detected in these 5-week-old LTC maintained a linear relationship to the number of input AML cells. Limiting dilution analysis, performed on 6 of the 10 samples, showed the frequency of AML cells initiating LTC (AML LTC-IC) to be 5- to 300-fold lower than the frequency of AML-CFC in the same cell sample, whereas the average number of CFC produced per LTC-IC varied from 1 to 13. Surprisingly, in each case the concentration of cytogenetically normal ITC-IC detected in AML patient blood was at least 10-fold higher than that previously observed in the blood of normal individuals. ''Mixed'' mouse fibroblast feeders engineered to produce human G-CSF,IL-3, and SF did not enhance detection of AML LTC-IC but did increase the output of cytogenetically normal CFC from LTC of 3 of 4 patient samples. Supplementation of AML LTC with IL-3 and exogenously provided SF and/or FL increased the output of AML-CFC from 5-week-old LTC by greater than or equal to twofold with 5 of 9 patient samples, whereas in one case exogenous addition of FL reduced the output of malignant CFC from LTC. These studies show that conditions that support normal LTC-IC also allow a functionally analogous but rare AML progenitor cell type to be detected. In addition, differences in the responses of normal and leukemic cells to various cytokines active on normal LTC-IC were revealed. Further analysis of these differences may enhance our understanding of leukemogenesis and lead to observations that could be exploited therapeutically. (C) 1997 by The American Society of Hematology.