流式细胞术对牛血中性粒细胞吞噬荧光细菌和酵母聚糖颗粒的特性分析
Flow cytometric characterization of bovine blood neutrophil phagocytosis of fluorescent bacteria and zymosan particles.
作者信息
Saad A M, Hageltorn M
出版信息
Acta Vet Scand. 1985;26(3):289-307. doi: 10.1186/BF03546530.
A Flow Cytometric method for the evaluation of the phagocytic capacity of bovine blood neutrophils is described. The neutrophils were isolated from bovine blood by a one step discontinuous gradient of Percoll. By this technique of isolation, 90 ± 2.8 % (mean ± s) of the granulocytes in the whole blood were recovered. Isolated neutrophils were incubated with FITC labeled S. aureus or zymosan particles in a ratio of 1:20 and 1:10, respectively, and a final serum concentration of 10 % Phagocytosis was terminated after 15 min and the number of extracellular bacteria or zymosan particles and the percentage of phagocytic granulocytes were registered by Flow Cytometry (FCM). FCM and microscopic studies revealed that eosinophils play a minor role in the phagocytosis of bacteria. The neutrophils were the main population of the granulocytes which were actively phagocytic. Variation among cows in the ability of their blood neutrophils to phagocytize bacteria was evident.
本文描述了一种用于评估牛血中性粒细胞吞噬能力的流式细胞术方法。通过一步不连续的Percoll梯度从牛血中分离出中性粒细胞。采用这种分离技术，全血中90±2.8%（平均值±标准差）的粒细胞得以回收。将分离出的中性粒细胞分别与异硫氰酸荧光素（FITC）标记的金黄色葡萄球菌或酵母聚糖颗粒以1:20和1:10的比例孵育，最终血清浓度为10%。15分钟后终止吞噬作用，通过流式细胞术（FCM）记录细胞外细菌或酵母聚糖颗粒的数量以及吞噬性粒细胞的百分比。FCM和显微镜研究表明，嗜酸性粒细胞在细菌吞噬中起次要作用。中性粒细胞是积极参与吞噬作用的粒细胞的主要群体。牛血中性粒细胞吞噬细菌的能力在不同奶牛之间存在明显差异。
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