van de Loosdrecht A A, Ossenkoppele G J, Beelen R H, Broekhoven M G, van Hooff M H, Dräger A M, Huijgens P C, Langenhuijsen M M
Department of Hematology, Academisch Ziekenhuis Vrije Universiteit, Amsterdam, The Netherlands.
Leukemia. 1994 Aug;8(8):1392-400.
In view of cellular immunotherapy with cytotoxic monocytes in minimal residual leukemia we have studied the effects of monocytes on the growth and survival of leukemic cells from cell lines and from patients with acute myeloid leukemia (AML). Using highly purified and interferon-gamma (IFN gamma) activated human monocytes, monocyte-mediated cytotoxicity (MMC) was evaluated in an MTT-based colorimetric cytotoxicity assay against six human leukemic cell lines (U937, THP1, KG1, K562, HL60, and 1,25(OH)2D3 differentiated HL60 cells) and cells from AML patients. Leukemic cells from cell lines with an immature phenotype were found to be resistant to MMC, whereas leukemic cells with a more mature and monocytic phenotype were sensitive. This paralleled the sensitivity to tumor necrosis factor-alpha (TNF-alpha). AML cells from patients with an immature phenotype (FAB-M1/M5A) were significantly less sensitive to MMC as compared to more mature AML cells (FAB-M2/M4/M5B). The growth stimulatory effects of non-activated monocytes on immature AML cells could be abrogated in the presence of IFN gamma or IL-3 and GM-CSF. In addition, these cytokines further potentiated MMC, preferentially affecting cells with a more mature phenotype. AML cells with an immunologically immature phenotype (CD34(high), HLA-Dr(low), CD13(low), CD14(low)) were revealed as the least sensitive cells to MMC. The growth stimulatory effects of IL-3/GM-CSF with or without TNF-alpha on AML cells correlated with resistance to MMC. In addition, the cytolytic effects of TNF-alpha in the presence of IFN gamma correlated with an increased susceptibility of AML cells to MMC. In conclusion, our data strongly indicate that MMC is related to maturation in AML, which is correlated to the differential stimulatory and cytolytic effects of monocyte-derived cytokines such as IL-3, GM-CSF, and TNF-alpha.
鉴于在微小残留白血病中采用细胞毒性单核细胞进行细胞免疫治疗，我们研究了单核细胞对白血病细胞系及急性髓系白血病（AML）患者白血病细胞生长和存活的影响。使用高度纯化且经干扰素-γ（IFNγ）激活的人单核细胞，通过基于MTT的比色细胞毒性测定法，评估单核细胞介导的细胞毒性（MMC）对六种人白血病细胞系（U937、THP1、KG1、K562、HL60以及1,25(OH)2D3分化的HL60细胞）和AML患者细胞的作用。发现具有未成熟表型的白血病细胞系对MMC有抗性，而具有更成熟和单核细胞表型的白血病细胞则敏感。这与对肿瘤坏死因子-α（TNF-α）的敏感性平行。与更成熟的AML细胞（FAB-M2/M4/M5B）相比，具有未成熟表型（FAB-M1/M5A）的AML患者细胞对MMC的敏感性明显较低。在IFNγ或IL-3和GM-CSF存在的情况下，未激活的单核细胞对未成熟AML细胞的生长刺激作用可被消除。此外，这些细胞因子进一步增强了MMC，优先影响具有更成熟表型的细胞。具有免疫未成熟表型（CD34(高)、HLA-Dr(低)、CD13(低)、CD14(低)）的AML细胞被发现是对MMC最不敏感的细胞。IL-3/GM-CSF无论有无TNF-α对AML细胞的生长刺激作用都与对MMC的抗性相关。此外，TNF-α在IFNγ存在下的溶细胞作用与AML细胞对MMC敏感性的增加相关。总之，我们的数据强烈表明MMC与AML中的成熟相关，这与单核细胞衍生的细胞因子如IL-3、GM-CSF和TNF-α的不同刺激和溶细胞作用相关。
