人红白血病细胞株K562培养上清对外周血单个核细胞的影响

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目的了解人红白血病细胞株K562分泌物对外周血单个核细胞(PBMC)的影响及其机制。方法选择K562细胞上清处理总的外周血单个核细胞(PBMC),首先用流式细胞术检测K562细胞上清对PBMC活化标志CD69和HLA-DR表达的影响,并通过改良的流式细胞术检测其增殖(数量)情况。然后进一步用流式细胞仪检测PBMC细胞的增殖指数和凋亡率以及各亚群细胞表面标志分子的表达情况。结果在K562细胞上清作用下,PBMC表面活化标志CD69和HLA-DR表达阳性细胞百分率显著增加,但是在K562细胞上清的影响下,PBMC数量明显下降。在检测对照组的增殖指数和凋亡率时,发现在培养72 h后,PBMC细胞的增殖指数明显上升,且凋亡率也显著增加。此外,在进一步对PBMC亚群的检测发现,与对照组相比,T细胞阳性细胞百分率明显下降,B细胞和单核细胞的百分率则显著上升,NK细胞无明显变化。CD4/CD8比值虽有所增加,但变化不明显。结论 K562上清虽能够诱导PBMC细胞活化,但也会诱导亚群中T细胞的大量凋亡,这可能是导致K562逃避免疫监视的原因之一。 Objective To investigate the effect of human erythroleukemia cell line K562 secretion on peripheral blood mononuclear cells (PBMCs) and its mechanism. Methods The total peripheral blood mononuclear cells (PBMCs) were treated with K562 cell supernatant. The effects of K562 cell supernatants on the expression of PBMC activation markers CD69 and HLA-DR were detected by flow cytometry. Test their proliferation (number) situation. Then the proliferation index and apoptosis rate of PBMCs and the expression of cell surface marker molecules of each subpopulation were further detected by flow cytometry. Results The percentage of PBMC positive cells expressing CD69 and HLA-DR was significantly increased under the action of K562 cell supernatant, but the number of PBMC decreased significantly under the influence of K562 cell supernatant. Proliferation index and apoptosis rate of control group were detected, and after 72 h of culture, the proliferation index of PBMC significantly increased, and the apoptosis rate also increased significantly. In addition, further detection of PBMC subpopulations found that the percentage of T-cell positive cells, the percentage of B cells and monocytes increased significantly compared with the control group, while the percentage of NK cells did not change significantly. Although CD4 / CD8 ratio increased, but the change is not obvious. Conclusion Although the supernatant of K562 can induce the activation of PBMC, it also induces the massive apoptosis of T cells in the subpopulation, which may be one of the reasons leading to the immune surveillance of K562.
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