自体树突状细胞用于慢性粒细胞白血病骨髓净化的初步研究

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 采用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（rhIL－4)、重组人粒－单集落刺激因子（rhGM－CSF）和重组人肿瘤坏死因子（rhTNFα）的RPMI－1640培养液。37℃CO2孵箱培养10天，或用免疫磁珠从PBMNC分离DC，在含自体血浆、rhGMCSF和rhTNFα的RPMI1640培养液培养3天，收获的细胞即DC加到含rhIL－2的Dexter骨髓长期培养体系，置37℃，5％CO2孵箱培养7天，观察前后骨髓细胞的免疫表型和P210阳性细胞比例。结果 从CML病人外周血获得的DC大多数表达P210。在含rhIL－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P210。在含rhIL－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P210阳性细胞减少，但在急性期病例，体系中P210细胞明显增加。结论 CML病人的DC表达P210，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。

Use of Autologous Dendritic Cells for Bone Marrow Purificationof Chronic Myelogenous Leukemia

Objective To investigate the effect of autologous dendritic cells(DC) to activate T lymphocytes and purify bone marrow of chronic myelogenous leukemia(CML). Methods Peripheral blood was collected from 3 cases of CML(2 in chronic phase and the other with acute transformation) and peripheral blood mononuclear cells(PBMNC) were seperated. DC were obtained by means of either culturing plastic-adherent PBMNC in RPMI 1640 containing autologous plasma, rhIL-4, rhGM-CSF and rhTNFα for 10 days or selecting negatively with immunological magnetic beads and culturing the selected cells in RPMI 1640 containing autologous plasma, rhGM-CSF and rhTNFα for 3 days. The obtained DC were added to the Dexter bone marrow long-term culture system with rhIL-2 and the culture was maintained for 7 days. Immunological phenotypes and percentage of P210 positive cells were analysed before and after the culture. Results To obtain DC from the peripheral blood of CML patients, the number were comparable to that of healthy volunteer, most of which express P210. Adding DC to Dexter system containing rhIL-2 appeard to further increase T cells in all three cases. In both cases with chronic phase, the percentage of P210 cells appeared to decrease. However, significant increase of P210 positive cells were found in the case with acute transformation. Conclusion Dendritic cells from CML patients do express P210 and are able to stimulate proliferation of T lymphocytes in the bone marrow. In chronic phase patients, DC-activated T lymphocytes appear to have some effects in purification of leukemic marrow.