Zheng Shui-er, Jin Jie, Tong Xiang-min
Department of Hematology, The First Affiliated Hospital, Zhejiang University Medical College, Hangzhou 310003, China.
Zhonghua Yi Xue Za Zhi. 2006 Aug 29;86(32):2252-5.
To investigate the effects of Imatinib mesylate (STI571) on the development of dendritic cells (DC) derived from the bone marrow mononuclear cells of patients with chronic myeloid leukemia (CML).
Bone marrow mononuclear cells (BMMNC) from CML patients were cultured initially using multiple cytokine combinations as follows: recombined human granulocyte/macrophage colony-stimulating-factor (rhGM-CSF) plus recombined human interleukin-4 (rhIL-4) as control groups, rhGM-CSF plus rhIL-4 and STI571 as experimental groups, and from day 8 added recombined human tumor necrosis factor-alpha (rhTNF-alpha) for stimulating maturation. The morphologic features of cells were observed by Wright's staining, Cytogenetic analysis was performed by Fluorescence in-situ hybridization (FISH), phenotypes were assessed by flow cytometry, and the functions of antigen-presenting were assayed by mixed lymphocyte reaction (MLR). The concentration of VEGF was detected by enzyme-linked immunosorbent assay (ELISA). NF-kappaB activation was evaluated by TransAM(TM) ELISA kit.
CML experimental groups treated with STI571 displayed features in morphology which were similar to those of control groups with delicate membrane projections. FISH confirmed the DC of both CML groups were leukemic origin. In comparison with the CML control groups, the CML experimental groups showed an increased expression of CD80, CD86, CD83 and HLA-DR and showed more intense abilities of allogeneic antigen presentation. The concentration of VEGF was dramatically reduced, and yet NF-kappaB activation was increased in experimental groups.
STI571 could promote the activation/maturation of DC derived from BMMNCs of patients with CML in vitro, which might be partially responsible for the fact that the inhibitory effect of VEGF on DC NF-kappaB activation was relieved through STI571 inhibiting the overproduction of VEGF in CML.
探讨甲磺酸伊马替尼（STI571）对慢性髓性白血病（CML）患者骨髓单个核细胞来源的树突状细胞（DC）发育的影响。
采用多种细胞因子组合对CML患者的骨髓单个核细胞（BMMNC）进行初始培养，具体如下：重组人粒细胞/巨噬细胞集落刺激因子（rhGM-CSF）加重组人白细胞介素-4（rhIL-4）作为对照组，rhGM-CSF加rhIL-4和STI571作为实验组，从第8天起添加重组人肿瘤坏死因子-α（rhTNF-α）以刺激成熟。通过瑞氏染色观察细胞形态特征，采用荧光原位杂交（FISH）进行细胞遗传学分析，通过流式细胞术评估细胞表型，并通过混合淋巴细胞反应（MLR）检测抗原呈递功能。采用酶联免疫吸附测定（ELISA）检测VEGF浓度。通过TransAM（TM）ELISA试剂盒评估NF-κB激活情况。
用STI571处理的CML实验组在形态上表现出与对照组相似的特征，有精细的膜突起。FISH证实两个CML组的DC均来源于白血病细胞。与CML对照组相比，CML实验组CD80、CD86、CD83和HLA-DR的表达增加，同种异体抗原呈递能力更强。VEGF浓度显著降低，而实验组中NF-κB激活增加。
STI571可在体外促进CML患者BMMNC来源的DC的激活/成熟，这可能部分解释了通过STI571抑制CML中VEGF的过度产生，VEGF对DC NF-κB激活的抑制作用得以解除这一现象。
