Hansel T T, Pound J D, Pilling D, Kitas G D, Salmon M, Gentle T A, Lee S S, Thompson R A
Regional Department of Immunology, East Birmingham Hospital, U.K.
J Immunol Methods. 1989 Aug 15;122(1):97-103. doi: 10.1016/0022-1759(89)90339-6.
A simple method for isolating highly purified eosinophils from human blood is described. Buffy coats from normal individuals (eosinophil counts less than 0.4 x 10(9)/litre) were centrifuged through a two layer Percoll density gradient, to produce a granulocyte fraction containing neutrophils and eosinophils. Neutrophils were extracted from this fraction using a monoclonal antibody (CLB FcR gran 1) against CD 16 (Fc gamma R III) in a direct or indirect selection procedure using immunomagnetic beads (Dynabeads). This negative immunoselection produced eosinophils of greater purity and with a superior capacity to mount a respiratory burst than eosinophils isolated by a method employing metrizamide.
本文描述了一种从人血中分离高纯度嗜酸性粒细胞的简单方法。取自正常个体(嗜酸性粒细胞计数低于0.4×10⁹/升)的血沉棕黄层通过两层Percoll密度梯度进行离心,以产生含有中性粒细胞和嗜酸性粒细胞的粒细胞部分。使用针对CD16(FcγR III)的单克隆抗体(CLB FcR gran 1),通过免疫磁珠(Dynabeads)的直接或间接选择程序,从该部分中提取中性粒细胞。这种阴性免疫选择产生的嗜酸性粒细胞比通过使用甲泛葡胺的方法分离的嗜酸性粒细胞具有更高的纯度和更强的产生呼吸爆发的能力。