流式细胞仪测定奶牛全血多形核白细胞的吞噬功能和呼吸爆发作用  

A Flow Cytometric Assay for Quantitating Phagocytic and Oxidative Burst Activity of Bovine Polymorphonuclear Leukocytes in Whole Blood

作　　者：胡松华[1] A.Johannisson 

机构地区：[1]浙江大学动物科学学院,浙江杭州310029 [2]瑞典农业大学兽医学院

出　　处：《中国兽医学报》2001年第6期586-588,共3页Chinese Journal of Veterinary Science

摘　　要：试验建立了用流式细胞仪同时测定奶牛全血中多形核白细胞 (PMN)吞噬功能和呼吸爆发作用的微量检测方法。奶牛全血用二氢诺丹明处理后 ,与直径为 1.75 μm的蓝色荧光乳胶微球培养 ,PMN内的二氢诺丹明被呼吸爆发过程中产生的还原性物质转化成发绿色荧光的诺丹明 (ROD)。用流式细胞仪计数 PMN,仪器可在数秒钟内计数几十万个 PMN,识别 PMN是否吞噬蓝色荧光微球和 /或含 ROD,并可检测每个 PMN所吞噬的微球数量。当血液中含细胞松弛素 B为 1、2 .5、5 mg/ L 时 。A rapid and sensitive flow cytometric method for simultaneous measurement of phagocytosis rate and oxidative burst activity of bovine polymorphonuclear leukocytes(PMN) in small amount of blood samples is described.After incubation of the PMN with 1.75 μm blue fluorescent beads,intracellular dihydrorhodamine 123(DHR) was oxidized into green fluorescent rhodamine 123(ROD) by oxidative burst products.Cells with beads and ROD were simultaneously detected by a flow cytometer.The method also allows to count the number of beads ingested by PMN.Phagocytosis and oxidative burst activity both increased with increasing beads concentration.A ratio of 50 beads to one cell was considered optimal for assaying bovine blood PMN activity.To evaluate the reliability of the method,blood samples were treated with a phagocytosis inhibitor,cytochalasin B.Incubation of blood with cytochalasin B at 1,2.5 or 5 mg/L resulted in a decrease in the phagocytosis rate with a concommitant decrease in oxidative burst activity.

关 键 词：奶牛 多形核白细胞 吞噬 呼吸爆发 流式细胞仪 

分 类 号：S858.23[农业科学—临床兽医学] S852.21[农业科学—兽医学]