流式细胞术检测外周血CD14^+细胞的活化程度

Application of flow cytometry to analyzing the activation states of peripheral blood mononuclear cells CD14 macrophage

目的 利用流式细胞术检测外周血CD14+细胞活化程度。方法 应用流式细胞术分别检测了 2 5份 ( 7份肝素抗凝的正常对照标本、7例肝素抗凝、11例枸橼酸钠抗凝的慢性活动性乙型肝炎标本 )外周血 (PBMC)CD14+细胞的侧向角光散射 (颗粒度 )SSC值、细胞活化相关表面抗原 (CD6 9)表达率。结果 肝素抗凝HBV组CD14+细胞颗粒度值 [( 85 3 1± 2 46 3)道 ]高于肝素抗凝正常对照组CD14+细胞颗粒度值 [( 474 5± 47 9)道 ](P≤ 0 0 5 ) ;肝素抗凝HBV组CD14+细胞颗粒度值 [( 85 3 1±2 46 3)道 ]高于枸橼酸钠抗凝组 [( 5 2 0 1± 10 5 6 )道 ](P≤ 0 0 5 ) ;肝素抗凝HBV组CD6 9在CD14+细胞的表达率 ( 2 2 71± 13 5 7) %明显高于肝素抗凝正常对照组CD6 9在CD14+细胞的表达率 ( 7 18±4 2 7) % (P≤ 0 0 5 ) ;肝素抗凝HBV组CD6 9在CD14+细胞的表达率 ( 2 2 71± 13 5 7) %明显高于枸橼酸钠抗凝HBV组 ( 3 93± 3 94) % (P≤ 0 0 1)。结论 采用流式细胞术检测细胞颗粒度、活化抗原表达率 ,是一种灵敏、快速、客观的评价外周血CD14+细胞活化程度的方法 ;使用不同抗凝剂肝素或枸橼酸钠对外周血CD14+细胞颗粒度、活化抗原表达率有影响 ,肝素抗凝血用来检测CD14+细胞活化程度效果更好。

Objective Analyzing the activation states of peripheral blood mononuclear cells PBMCs CD14 macrophage by flow cytometry (FCM). Method 7 heparin treated blood and 11 Tri-Sodium Citrate treated blood were collected from chronic active hepatitis patients, SSC (granulate) and activation-linked surface antigen CD69 expression rate of CD14 cells were tested by FCM. Results The SSC (granulate) mean value of heparin treated blood, CD14 cells (853.1±246.3channel) was high than that of health control [(474.5±47.9)channel] (P≤0.05); The SSC (granulate) mean value of heparin treated blood, CD14 cells [(853.1±246.3) channel] was high than that of Tri-Sodium Citrate treated blood [(520.1±105.6) channel] (P≤0.05); heparinized blood activation-linked surface antigen CD69 expression rate of CD14 cells (22.71±13.57) % was high than that of health control (7.18±4.27) % (P≤0.05); heparin treated blood activation-linked surface antigen CD69 expression rate of CD14 cells (22.71±13.57) % was significant high than that of Tri-Sodium Citrate treated blood (3.93±3.94)% (P≤0.01). The SSC (granulate) mean value of HBV CD14 cells has no relevance with serum ALT level, there was significant correlation between SSC (granulate) mean value and liver cirrhosis; Statistic analysis showed no correlation between CD14 cells CD69 expression rate and serum ALT level or liver cirrhosis. Conclusion FCM measurement may be characterized by high sensitivity, rapidity, and objectivity for the determination of PBMC CD14 macrophage activation states. The granulate and activation-linked surface antigen CD69 expression rate of CD14 cells were differed in Tri-Sodium Citrate or heparin-treated blood, and CD14 cell activation was better obtained in heparinized blood.