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一种纯化人嗜酸性粒细胞和中性粒细胞的新方法,以及这些细胞损伤曼氏血吸虫童虫能力的比较。

A new method for the purification of human eosinophils and neutrophils, and a comparison of the ability of these cells to damage schistosomula of Schistosoma mansoni.

作者信息

Vadas M A, David J R, Butterworth A, Pisani N T, Siongok T A

出版信息

J Immunol. 1979 Apr;122(4):1228-36.

PMID:448088
Abstract

Centrifugation of human white blood cells over either Ficoll-Hypaque or slightly hypertonic Metrizamide discontinuous gradients reliably produces separate fractions that are enriched for either neutrophilic or eosinophilic granulocytes. This single step purification routinely yields 90 to 100% pure neutrophils and 85 to 100% pure eosinophils. Metrizamide gradients, in particular, reproducibly provide high yields of 90 to 100% pure eosinophils from normal subjects with 2 to 3% eosinophils in their peripheral blood. The method does not damage cells as judged by morphologic or functional criteria. The purified cell populations were tested for their ability to damage antibody-coated schistosomula either by the measurement of 51Cr release from labeled organisms, or by direct morphologic assessment. Neutrophils were superior in their ability to release 51Cr from labeled organisms, but eosinophils adhered to the organisms to a greater extent and induced microscopically detectable damage.

摘要

将人白细胞置于Ficoll-泛影葡胺或略高渗的甲泛葡胺不连续梯度上进行离心,可可靠地产生分别富含嗜中性粒细胞或嗜酸性粒细胞的分离组分。这一步骤的纯化常规可获得90%至100%纯度的嗜中性粒细胞和85%至100%纯度的嗜酸性粒细胞。特别是甲泛葡胺梯度,可重复性地从外周血嗜酸性粒细胞占2%至3%的正常受试者中获得90%至100%纯度的高产嗜酸性粒细胞。根据形态学或功能标准判断,该方法不会损伤细胞。通过测量标记生物体中51Cr的释放量,或通过直接形态学评估,对纯化的细胞群体损伤抗体包被的血吸虫幼虫的能力进行了测试。嗜中性粒细胞从标记生物体中释放51Cr的能力更强,但嗜酸性粒细胞与生物体的粘附程度更高,并能诱导显微镜下可检测到的损伤。

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