Korycka Anna, Robak Tadeusz
Department of Hematology, Medical University of Lódź, Pabianicka 62, 93-513 Lódź, Poland.
Arch Immunol Ther Exp (Warsz). 2003;51(1):61-7.
We evaluated the influence of cyclosporin A (CsA) used alone or together with the new purine nucleoside analogues (PNAs) 2-chlorodeoxyadenosine (2-CdA) and fludarabine (F-ara-A) on the colony growth of normal and chronic myelogenous leukemia (CML) granulocyte-macrophage progenitor cells (CFU-GM) in cultures in vitro. The assay was based on the Iscove's method in our modification. Specimens of bone marrow were collected from 15 patients with CML in the chronic phase and from 10 hematologically normal patients. CsA at the concentrations of 1, 2 and 4 microg/ml was used alone and, at the concentration of 4 microg/ml, was preincubated with mononuclear cells (MNCs) and, after 30 min PNAs were added to the culture medium. Concentrations of 5, 10, and 20 nM 2-CdA and 0.4, 0.8 and 1.6 microM F-ara-A were used. After 14 days of incubation, the colonies were scored under an inverted microscope. We observed that CsA used alone at all three concentrations inhibited the colony growth of CML CFU-GM to a statistically significant degree compared with the control (p < 0.02) and that it did not significantly influence normal colony growth. The IC50 for CsA was 3.9 microg/ml in the case of normal CFU-GM and 2.7 microg/ml in the case of CML CFU-GM. After the use of CsA in combination with either the highest concentrations of 2-CdA or F-ara-A, statistically significant differences compared with CsA used alone were observed (p = 0.008, p = 0.03 for CsA with 2-CdA, and p = 0.0007, p = 0.005 for CsA with F-ara-A, respectively, for normal and CML CFU-GM). However, there were no significant differences between the combinations of drugs and PNAs used alone. In the case of the combination of CsA with the highest concentrations of both PNAs, significant differences in colony growth inhibition between normal and CML CFU-GM were observed (p = 0.002 and p = 0.005, respectively, for 2-CdA and F-ara-A). In conclusion, at the concentrations of the drugs used, a subadditive action was observed either between CsA and 2-CdA or between CsA and F-ara-A.
我们评估了单独使用环孢素A（CsA）或其与新型嘌呤核苷类似物（PNA）2-氯脱氧腺苷（2-CdA）和氟达拉滨（F-ara-A）联合使用时，对体外培养的正常及慢性髓性白血病（CML）粒细胞-巨噬细胞祖细胞（CFU-GM）集落生长的影响。该实验基于我们改良的伊斯科夫方法。从15例慢性期CML患者及10例血液学正常患者采集骨髓样本。分别单独使用浓度为1、2和4μg/ml的CsA，并在浓度为4μg/ml时与单核细胞（MNC）预孵育30分钟，之后向培养基中加入PNA。使用的2-CdA浓度为5、10和20 nM，F-ara-A浓度为0.4、0.8和1.6μM。孵育14天后，在倒置显微镜下对集落进行计数。我们观察到，与对照组相比，单独使用的所有三种浓度的CsA均能显著抑制CML CFU-GM的集落生长（p < 0.02），且对正常集落生长无显著影响。正常CFU-GM的CsA半数抑制浓度（IC50）为3.9μg/ml，CML CFU-GM的为2.7μg/ml。在将CsA与最高浓度的2-CdA或F-ara-A联合使用后，与单独使用CsA相比，观察到有统计学显著差异（对于正常及CML CFU-GM，CsA与2-CdA联合使用时p = 0.008，p = 0.03；CsA与F-ara-A联合使用时p = 0.0007，p = 0.005）。然而，药物联合使用与单独使用PNA之间无显著差异。在CsA与两种PNA的最高浓度联合使用的情况下，观察到正常及CML CFU-GM在集落生长抑制方面存在显著差异（对于2-CdA和F-ara-A，分别为p = 0.002和p = 0.005）。总之，在所使用的药物浓度下，观察到CsA与2-CdA之间或CsA与F-ara-A之间存在次加性作用。
