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目的 考察慢性粒细胞白血病 (CML)树突状细胞 (DC)刺激自体骨髓 T细胞 ,清除白血病细胞的作用。 方法 采集 3例 CML 病人 (2例慢性期和 1例急变期 )外周血 ,分离单个核细胞 (PBMNC) ,经塑料贴壁后在含自体血浆、重组人白介素 4(rh IL- 4)、重组人粒 -单集落刺激因子 (rh GM- CSF)和重组人肿瘤坏死因子 (rh TNFα)的 RPMI- 16 40培养液 ,37℃ ,5 % CO2 孵箱培养 10天 ,或用免疫磁珠从 PBMNC分离 DC,在含自体血浆、rh GM-CSF和 rh TNFα的 RPMI16 40培养液培养 3天 ,收获的细胞即 DC加到含 rh IL- 2的 Dexter骨髓长期培养体系 ,置37℃ ,5 % CO2 孵箱培养 7天 ,观察培养前后骨髓细胞的免疫表型和 P2 10阳性细胞比例。 结果 从 CML 病人外周血获得的 DC大多数表达 P2 10。在含 rh IL- 2的 Dexter体系加入 DC能进一步增加体系中的 T细胞数。在 2例慢性期病人体系中的 P2 10阳性细胞减少 ,但在急性期病例 ,体系中 P2 10细胞明显增加。 结论 CML 病人的 DC表达 P2 10 ,能刺激自体骨髓中 T细胞增生 ,在慢性期病人 ,这些自体 DC活化的骨髓 T细胞有清除白血病细胞的作用
Objective To investigate the effect of chronic myelogenous leukemia (CML) dendritic cells (DCs) on the stimulation of autologous bone marrow T cells to clear leukemia cells. Methods Peripheral blood was collected from 3 patients with CML (2 chronic phases and 1 blast crisis phase) and mononuclear cells (PBMNC) were isolated. After plastic adherence, autologous plasma, recombinant human interleukin 4 (rhIL-4), and recombination were performed. Human granulocyte-single colony stimulating factor (rh GM-CSF) and recombinant human tumor necrosis factor (rh TNFα) in RPMI-16 40 medium, 37°C, 5 % CO2 incubator for 10 days, or with immunomagnetic beads from PBMNC DCs were isolated and cultured in RPMI16 40 medium containing autologous plasma, rh GM-CSF and rh TNFα for 3 days. The harvested cells were added to a Dexter-containing bone marrow long-term culture system containing rh IL-2 at 37°C, 5% CO2. After incubation for 7 days in the incubator, the immunophenotype and P2 10 positive cell ratio of the bone marrow cells before and after culture were observed. Results Most DCs obtained from peripheral blood of CML patients expressed P2 10 . The addition of DC to the Dexter system containing rhIL-2 can further increase the number of T cells in the system. The number of P2 10 positive cells in the two chronic phase patient systems decreased, but in the acute phase, the P2 10 cells increased significantly. Conclusion The expression of P2 10 in DC of CML patients can stimulate the proliferation of T cells in autologous bone marrow. In the chronic phase, these autologous DC-activated bone marrow T cells have the function of eliminating leukemia cells.