细胞因子在体外对骨髓单个核细胞形态和免疫表型的影响

Morphology and Immunophenotype Study of Bone Marrow Mononuclear Cells after Activated by Different Cytokines In Vitro

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 cells in cultures were divided into four groups and cultured with various cytokines in vitro. These four groups are: control, IL-2 group, CD3-AK group, and CIK group. The morphological (cell volume, nucleus/plasm) changes of bone marrow cells in culture were observed. Immunophenotype analysis(CD34, CD38, CD3, CD56) were done before and after culture in all groups. Cytotoxicity against fresh acute leukemia cells were detected by modified MTT methods. The cell volume became larger with increased nucleus/plasm ratio in IL-2 group, CD3-AK group and CIK group. The plasm filled with PAS positive granules in most of cells in CD3-AK group and CIK group. The positive ratio of CD3, CD56, CD38 in CD3-AK or CIK group increased markedly after culture(P<0.05), but no significant difference between the two groups. The CD56 + cell increased in IL-2 group. CD34 + cells decreased in all groups and there were no significant differences among those four groups. The cytotoxicity to fresh leukemia cells: CD3-AK group and CIK group >IL-2 group >control group. There was no significant difference between CD3-AK group and CIK group. This experiment showed different effect on bone marrow cells by different cytokine combination. The cytokine combination of CD3-AK group or CIK group can make immunocytes of bone marrow proliferating and retained certain amount of stem/progenitor cells.