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目的 :利用自然杀伤细胞 (NK)的早期活化标记CD69的表达探讨NK细胞体外活化的规律 ,促进这类细胞的临床应用 方法 :用Ficoll-Hypaque梯度离心法分离出正常成人外周血单个核细胞 (PBMC) ,分别在两种不同的有丝分裂原 ,植物血凝素 (PHA ,2 0 μg/mL)和佛波醇酯 (PDB ,10 - 7mol/L)存在的条件下进行培养 ,于 0、2、12、2 4h收获细胞后进行双色免疫荧光标记 ,以流式细胞术对NK细胞的CD69分子表达情况进行分析 结果 :NK细胞在PHA、PDB刺激 2h后CD69的表达明显上升 ,而且随时间的延长CD69的表达率逐渐增加 ,2 4h分别达到 ( 84 .96± 9.2 4 ) %、( 91 85± 2 .94 ) % ;PDB刺激组CD69的表达率在不同的时间点均比PHA组高 结论 :体外条件下 ,PHA、PDB均能快速上调NK细胞CD69表达 ,而且PDB( 10 - 7mol/L)作用明显强于PHA( 2 0 μg/mL)
OBJECTIVE: To study the regulation of in vitro activation of natural killer (NK) NK cells in vitro and to promote the clinical application of NK cells. Methods: Normal adult peripheral blood mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation PBMC) were cultured in the presence of two different mitogen, phytohemagglutinin (PHA, 20 μg / mL) and phorbol ester (PDB, 10-7 mol / L) , 12,2 4h after harvesting cells were immunofluorescent labeling, expression of CD69 molecules in NK cells by flow cytometry results: NK cells in PHA, PDB stimulation 2h after CD69 expression increased significantly, and with the passage of time The expression of CD69 in PDB stimulation group was significantly higher than that in PHA group at 24 h (84.96 ± 9.24)% and (91 85 ± 2. 94)%, respectively In vitro, PHA and PDB could up - regulate the expression of CD69 in NK cells, and the effect of PDB (10 - 7mol / L) was stronger than that of PHA (20μg / mL)