Hansel T T, De Vries I J, Iff T, Rihs S, Wandzilak M, Betz S, Blaser K, Walker C
Swiss Institute of Allergy and Asthma Research (SIAF), Davos-Platz.
J Immunol Methods. 1991 Dec 15;145(1-2):105-10. doi: 10.1016/0022-1759(91)90315-7.
A simple method is described for the isolation of human peripheral blood eosinophils by an immunomagnetic procedure. Superparamagnetic particles were coupled to a monoclonal antibody against CD16, a molecule present on neutrophils but not on eosinophils. A peripheral blood granulocyte preparation, containing neutrophils and eosinophils, was incubated with these anti-CD16 particles. In the magnetic field of a permanent magnet, magnetically labelled neutrophils were then retained on columns with a ferromagnetic matrix. By this negative selection procedure, eosinophils of 99.5% purity were obtained from normal individuals and 99.6% purity from patients with eczema. A comparison was made between the immunomagnetic method and eosinophil isolation after N-formyl-methionyl-leucyl-phenyl-alanine treatment. Even in the case of individuals with very low eosinophil counts, the immunomagnetic method permits the efficient isolation of highly purified and functionally active eosinophils.
描述了一种通过免疫磁选法分离人外周血嗜酸性粒细胞的简单方法。超顺磁性颗粒与抗CD16单克隆抗体偶联，CD16是一种存在于中性粒细胞而非嗜酸性粒细胞上的分子。将含有中性粒细胞和嗜酸性粒细胞的外周血粒细胞制剂与这些抗CD16颗粒一起孵育。在永久磁铁的磁场中，磁性标记的中性粒细胞随后被保留在具有铁磁基质的柱上。通过这种阴性选择程序，从正常个体中获得了纯度为99.5%的嗜酸性粒细胞，从湿疹患者中获得了纯度为99.6%的嗜酸性粒细胞。对免疫磁选法与经N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸处理后的嗜酸性粒细胞分离方法进行了比较。即使在嗜酸性粒细胞计数非常低的个体中，免疫磁选法也能有效分离出高度纯化且功能活跃的嗜酸性粒细胞。
