论文部分内容阅读
为考察慢性粒细胞白血病 (CML)患者自体树突状细胞 (DCs)活化骨髓细胞、净化骨髓的作用 ,采用免疫磁珠从 2例血液学缓解的CML骨髓单个核细胞 (BMMNCs)分离DCs,另取BMMNCs置T 2 5塑料培养瓶建立Dexter培养体系 ,分为 3份 ,第 1份为对照 ,第 2份加rhIL 2 ,第3份加自体DCs。每周取半量非贴壁细胞、计数、行CFU GM检测 ,然后加等量新鲜培养基继续培养。当非贴壁细胞总数 <2× 10 5时终止培养 ,收集贴壁细胞做CFU GM集落形成试验并检测CD34和P2 10表达。取贴壁细胞形成的CFU GM集落 ,抽提RNA ,用反转录聚合酶链反应 (RT PCR)检测BCR ABL的表达。结果发现 ,在DCs加入CML骨髓Dexter培养体系培养后 1- 2周 ,非贴壁细胞的CFU GM产量明显下降 ,与rhIL 2的体系基本平行 ,同时体系中的P2 10阳性细胞显著减少。不过 ,培养 3周后含DCs的体系CFU GM产量下降减缓 ,而含rhIL 2的体系CFU GM产量持续下降。在含自体DCs的Dexter体系 ,贴壁细胞中CD34+ 细胞和P2 10 + 细胞比例最低 ,只是贴壁细胞产生的CFU GM集落中 ,BCR ABL(+ )的集落比例与不含DCs的体系比较无明显差别。这些结果说明自体DCs加入CML骨髓Dexter培养体系能减少白血病的祖细胞 ,包括成熟的祖细胞和原始的祖细胞 ,自体DCs有可能用于CML骨髓的体外净化。
To investigate the role of autologous dendritic cells (DCs) in activating myeloid cells and purifying bone marrow in patients with chronic myelocytic leukemia (CML), we used immunomagnetic beads to separate DCs from two hematologically resorbed CML bone marrow mononuclear cells (BMMNCs). BMMNCs were placed in T 2 5 plastic culture flasks to establish a Dexter culture system. The culture was divided into 3 groups. The first was control, the second was rhIL 2 , and the third was autologous DCs. Take half a day of non-adherent cells, count, and perform CFU GM test, then add equal amount of fresh medium to continue the culture. When the total number of non-adherent cells < 2 × 10 5 was terminated, the adherent cells were collected for CFU GM colony formation assay and tested for CD34 and P2 10 expression. CFU GM colonies formed by adherent cells were taken, RNA was extracted, and BCR ABL expression was detected by reverse transcription polymerase chain reaction (RT PCR). The results showed that after DCs were added to CML bone marrow Dexter culture system for 1 to 2 weeks, the CFU GM production of non-adherent cells decreased significantly, and the system was basically parallel to the rhIL 2 system, and the number of P2 10 positive cells in the system was significantly reduced. However, the production of CFU GM in the system containing DCs decreased after 3 weeks of cultivation, while the production of CFU GM in the system containing rhIL 2 continued to decline. In the Dexter system with autologous DCs, the proportion of CD34+ cells and P2 10+ cells in the adherent cells was the lowest, but only in the CFU GM colonies produced by the adherent cells. The proportion of BCR ABL(+) colonies was not significantly different from the system without DCs. difference. These results suggest that the addition of autologous DCs to CML bone marrow Dexter culture system can reduce leukemic progenitor cells, including mature progenitor cells and primitive progenitor cells. Autologous DCs may be used for the in vitro purification of CML bone marrow.