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树突状细胞(dendritic cells,DC)在激发机体抗白血病T细胞免疫应答中具有重要作用,有报道DC可以由单核细胞及粒细胞分化生成,本研究应用细胞因子在体外诱导了急性早幼粒细胞性白血病(APL)细胞向DC的分化.从M3型APL患者外周血分离白血病细胞,加入GM-CSF(100ng/ml)或GM-CSF(100ng/ml)+rhIL-4(500U/ml)体外培养14天,并于培养结束前3天加人TN-α(100ng/ml).结果表明,GM-CSF可以促进白血病细胞体外增殖,并从幼稚状态逐渐分化成熟,表达高水平的CD45分子,其中部分为CD14~+的单核细胞,部分为CDla~+的DC(M3DC);培养后期加入TNF-α,可以促进DC生成,约占35%;以GM-CSF+IL-4培养也诱导了幼稚细胞的成熟,2周后DC约占10%,但较少单核细胞生成,培养至3周时DC约占60%;而在培养后第11天加入TNF-α则可以加速DC生成,3天后生成的白血病型DC达90%.电镜观察到M3DC具有与单核细胞来源的DC相似的超微结构,但具有的突出特征是部分细胞存在少量胞浆颗粒;M3DC高表达HLA-DR、B7-2、CD40、CD54分子,体外可以强烈刺激同种T细胞增殖.此类由白血病细胞诱导生成的DC可被用于体内外诱导生成肿瘤特异性CTL,在APL的免疫治疗中具有一定的应用价值.
Dendritic cells (DCs) play an important role in stimulating the body’s immune response against leukemic T cells. DCs have been reported to be differentiated from monocytes and granulocytes. In this study, cytokines were used to induce acute early childhood Differentiation of granulocytic leukemia (APL) cells into DCs. Leukemia cells were isolated from the peripheral blood of M3 APL patients and added with GM-CSF (100 ng/ml) or GM-CSF (100 ng/ml) + rhIL-4 (500 U/ml ) Cultured in vitro for 14 days and added TN-α (100 ng/ml) 3 days before the end of culture. The results show that GM-CSF can promote the proliferation of leukemic cells in vitro and gradually differentiate and mature from the naive state, expressing high levels of CD45 Molecules, some of which are CD14~+ monocytes, some of which are CDla~+ DCs (M3DCs); the addition of TNF-a at the later stage of culture can promote DC production, accounting for about 35%; cultured with GM-CSF+IL-4. The maturation of naive cells was also induced. DCs accounted for approximately 10% after 2 weeks, but less monocytes were generated. DCs accounted for approximately 60% of cultures at 3 weeks; however, TNF-α was accelerated on the 11th day after culture. After DC generation, 90% of the leukemic DCs were generated after 3 days. Electron microscopy observed that the M3DCs had an ultrastructure similar to that of monocyte-derived DCs but had prominent features. There is a small amount of cytoplasmic granules in some cells; M3DC highly expresses HLA-DR, B7-2, CD40, and CD54 molecules and can strongly stimulate the proliferation of allogeneic T cells in vitro. Such DCs induced by leukemic cells can be used both in vitro and in vivo. The induction of the generation of tumor-specific CTL has a certain application value in the immunotherapy of APL.