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通过加入不同细胞因子 ,将骨髓单个核细胞分为对照组、IL 2组、CD3 AK组和CIK组进行体外培养。在培养过程中观察各组细胞体积、细胞核、细胞浆和细胞化学染色 (PAS ,POX)等细胞形态学的变化 ;培养前后各组取样以检测细胞免疫表型CD3,CD5 6,CD34和CD38;用改良的MTT法检测各组培养后对新鲜急性白血病细胞的杀伤情况。研究发现 :在培养过程中IL 2组、CD3 AK组和CIK组细胞体积、胞核增大 ,胞浆增多 ,胞浆中出现PAS阳性颗粒 ,培养后在CD3 AK组和CIK组大颗粒样幼稚淋巴细胞明显增加 ,且还可见较多PAS和POX阳性的单核 巨噬样大细胞 ;CD3 AK和CIK组CD3+ ,CD5 6+ ,CD38+ 细胞较培养前明显增加 (P <0 .0 5 ) ,但两组间无明显差别 ;IL 2组在培养后CD5 6+ 细胞也有增加 ,但CD3+ 和CD38+ 细胞无明显增加 ;各组培养后CD34+ 细胞比例均下降 (P <0 .0 5 ) ,但各组间无显著性差异 (P >0 .0 5 ) ;IL 2组、CD3 AK组和CIK组对新鲜急性白血病细胞的杀伤作用显著强于对照组 (P <0 .0 5 ) ,CD3 AK组、CIK组显著强于IL 2组 (P <0 .0 5 ) ,CD3 AK组与CIK组间无明显差异 (P >0 .0 5 )。本实验提示不同细胞因子组合对骨髓细胞的作用效果不同 ,CD3 AK组和CIK组的细胞因子组合既能刺激骨髓免疫细胞增殖和细胞毒增加 ,又能保留一定量?
Bone marrow mononuclear cells were divided into control group, IL 2 group, CD3 AK group and CIK group by adding different cytokines for in vitro culture. The morphological changes of cells such as cell volume, nucleus, cytoplasm and cytochemical staining (PAS, POX) were observed during the culture. Before and after culturing, the immunophenotypes of CD3, CD5 6, CD34 and CD38 were detected. The modified MTT assay was used to detect the killing of fresh acute leukemia cells in each group. The results showed that the volume of cells, nucleus and cytoplasm increased in IL 2, CD3 AK and CIK groups during culture, and PAS positive particles appeared in cytoplasm. After culturing, there were large particles in CD3 AK group and CIK group Lymphocytes were significantly increased, and more PAS and POX positive macrophage macrophage-like macrophages were also observed. CD3 +, CD5 + and CD38 + cells in CD3-AK and CIK groups were significantly increased compared with that before culture (P <0.05) However, there was no significant difference between the two groups. CD5 6+ cells also increased in IL2 group, but there was no significant increase in CD3 + and CD38 + cells. The percentage of CD34 + cells in each group decreased (P <0.05) (P> 0.05). The killing effect of IL 2, CD3 AK and CIK on fresh acute leukemia cells was significantly stronger than that of the control group (P <0.05), and the CD3 AK group , CIK group was significantly stronger than IL 2 group (P <0.05), CD3 AK group and CIK group no significant difference (P> 0.05). This experiment suggests that different combinations of cytokines on bone marrow cells effect is different, CD3 AK group and CIK group of cytokine combinations can both stimulate bone marrow immune cell proliferation and cytotoxicity, but also retain a certain amount?