Optimizing preparation of normal dendritic cells and bcr-abl+ mature dendritic cells derived from immunomagnetically purified CD14+ cells
Optimizing preparation of normal dendritic cells and bcr-abl+ mature dendritic cells derived from immunomagnetically purified CD14+ cells
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DOI：
10.1089/152581600319676
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发表时间：
2000-02-01
期刊：
影响因子：
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通讯作者：
Vuk-Pavlovic, S
中科院分区：
文献类型：
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作者：
Dietz, AB;Bulur, PA;Vuk-Pavlovic, S
本研究的目的是优化从慢性髓性白血病（CML）患者中获得的树突状细胞（DC）制剂，并将其与从正常CD 14（+）单核细胞（MNC）中制备的DC进行比较。我们研究了正常DC和bcr-abl（+）白血病DC（CML-DC）的产量、膜分子表达、分化状态和刺激T细胞的能力。我们通过CD 14特异性免疫吸附从PBMC中分离DC前体，并将其在GM-CSF和IL-4中培养7天，然后孵育3天以使细胞完全分化。我们使用补充有FBS的RPMI 1640培养基和含有人AB血清的X-VIVO 15培养基评估CML-DC的培养物。与在RPMI 1640中成熟的细胞相反，几乎所有在X-VIVO 15中孵育的细胞都表达成熟DC的标志物CD 83。CML-DC和正常DC在CD 83的表达上是不可区分的，导致迄今为止报道的最高百分比。CD 14（+）细胞诱导的正常DC和CML-DC产量无明显差异。PBMC中bcr-abl（+）细胞的百分比在患者中在65%和97%之间变化，并且最终的CML-DC制备物为>98% bcr-abl（+），这是迄今为止bcr-abl（+）细胞的最高纯度。正常DC和CML-DC在刺激同种异体和自体T细胞增殖方面同样有效。这些技术提供了高度富集的、成熟的、功能性的CML-DC。
The goal of this work was to optimize dendritic cell (DC) preparations obtained from patients suffering from chronic myeloid leukemia (CML) and compare them with DC prepared from normal CD14(+) mononuclear cells (MNC). We studied normal DC and bcr-abl(+) leukemic DC (CML-DC) yields, expression of membrane molecules, differentiation status, and ability to stimulate T cells. We isolated DC precursors from PBMC by CD14-specific immunoadsorption and cultured them for 7 days in GM-CSF and IL-4, followed by a 3-day incubation to fully differentiate the cells. We evaluated cultures of CML-DC using RPMI 1640 medium supplemented with FBS and X-VIVO 15 medium containing human AB serum. In contrast to cells matured in RPMI 1640, virtually all cells incubated in X-VIVO 15 expressed CD83, a marker of mature DC. CML-DC and normal DC were indistinguishable in expression of CD83, resulting in the highest percentage reported so far. The yields of normal DC and CML-DC from CD14(+) cells were indistinguishable. The percentage of bcr-abl(+) cells in PBMC varied among patients between 65% and 97% and the final CML-DC preparations were >98% bcr-abl(+) the highest purity of bcr-abl(+) cells to date. Normal DC and CML-DC were equally effective in stimulating proliferation of allogeneic and autologous T cells. These techniques provide highly enriched, mature, functional CML-DC.