外周血巨噬细胞功能状态检测方法的建立

Establishment of method to determine function of PBMC macrophage

目的 运用荧光标记的抗细胞表面抗原和抗细胞因子抗原 ,结合固定、破膜处理技术 ,建立四色荧光染色法流式细胞术检测巨噬细胞功能状态的方法。方法 应用流式细胞术检测了 1例肝素正常对照 ,1例肝素抗凝的慢性活动性乙型肝炎外周血 (PBMC)CD14 + 巨噬细胞的吞噬率以及前向角光散射 (细胞大小 )FSC值、侧向角光散射 (颗粒度 )SSC值、细胞活化相关表面抗原 (CD6 9)、核细胞增殖相关抗原 (Ki 6 7)、肿瘤坏死因子 (TNF a)的表达率。结果 SSC、吞噬率、CD6 9、Ki 6 7和TNF a表达率在同一样本表达水平一致。结论 采用流式细胞术检测细胞颗粒度、吞噬率、Ki 6 7和TNF a的表达率 ,是又一灵敏、快速、客观评价外周血CD14 +

Objective: Using labeled cell surface antigen antibodies and anti cytokine antibodies with a combination of fixation and permeabilization method, we have established the method for measuring function stages of activated macrophage of peripheral blood mononuclear cells (PBMC) by 4 colors flow cytometry (FCM) Method: 1 heparin sodium treated blood from volunteer and 1 heparin sodium treated blood from chronic active hepatitis patient were collected, FSC(forward scatter, size)?SSC(side scatter, granulate)?phagocytosis rates?activation linked surface antigen CD69?nuclear cell proliferation associated antigen(Ki 67)and intracellular cytokines tumor necrosis factor(TNF a) expression rates of CD14+ cells were tested by FCM Results: SSC?phagocytosis rates?CD69?Ki 67 and TNF a rates were consistent in the same sample Conclusion: FCM measurement may be characterized as a high sensitive, rapid, and objective method for determining the function states of activated PBMC CD14 + macrophage