甲磺酸伊马替尼对慢性髓细胞白血病树突状细胞发育的影响及机制探讨

Effects of Imatinib mesyiate on the development of dendritic cells derived from bone marrow mononuclear cells of patients with chronic myeloid leukemia

目的研究甲磺酸伊马替尼(Imatinib,STI571)对慢性髓细胞白血病(CML)树突状细胞(DC)发育的影响及探讨其可能作用机制。方法分离CML骨髓单个核细胞,在培养体系中加入重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)、重组人白细胞介素4(rhIL-4)培养(对照组)和在此基础上再加入STI571培养(实验组),于第8天加入重组人肿瘤坏死因子α(rhTNF-α)进一步刺激成熟。观察培养后细胞形态、表型、混合淋巴细胞反应等指标,检测培养上清血管内皮生长因子(VEGF)浓度、细胞核因子κB(NF-κB)活性。结果2组均呈现典型的树突状细胞形态,实验组CD80、CD86、CD83和人类白细胞抗原DR(HLA-DR)的表达、刺激淋巴细胞增殖的能力显著高于对照组(均P<0．05);实验组VEGF浓度显著低于对照组(P<0．05),而NF-κB活性显著高于对照组(P< 0．05)。结论STI571可促进CML骨髓来源DC的活化,这可能与其解除了CML来源VEGF对DC NF-κB活性的抑制有关。

Objective To investigated 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）. Methods 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-α （rhTNF-α） 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-KB activation was evaluated by TransAMTM ELISA kit. Results CML experimental groups treated with STL571 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-KB activation was increased in experimental groups. Conclusion STI571 could promoted 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-KB activation was relieved through STL571 inhibiting the overproduction of VEGF in CML.