Landry F J, Findlay S R
J Immunol Methods. 1983 Oct 28;63(3):329-36. doi: 10.1016/s0022-1759(83)80006-4.
Enriched populations of human basophils were prepared using a combination of negative selection techniques. First, a 2-step discontinuous gradient was made using 65% and 55% isotonic Percoll. Purification of plasma basophils by centrifugation through the gradient increased basophils from 0.96 +/- 0.55% to 14.6 +/- 7.9% (n = 9). In 5 other experiments a second step was added in which contaminating mononuclear cells were removed using a panning technique. In this technique, Percoll separated cells were treated with anti-T-cell antibodies. Contaminating T-cells were selectively removed by adherence to a petri dish coated with affinity purified goat anti-mouse IgG. Basophil purity was increased to 34 +/- 15% using this step. These experiments demonstrated that negative selection techniques can yield an increase in basophil purity 50-100-fold. The purified basophils contained the expected quantity of histamine (1.36-1.54 pg/basophil) and released the same amount of histamine in response response to different concentrations of goat anti-human IgE Fc as did unpurified cells.
使用多种阴性选择技术制备富集的人嗜碱性粒细胞群体。首先，使用65%和55%的等渗 Percoll 制成两步不连续梯度。通过该梯度离心纯化血浆嗜碱性粒细胞，使嗜碱性粒细胞比例从0.96±0.55%增加到14.6±7.9%（n = 9）。在另外5个实验中增加了第二步，即使用淘选技术去除污染的单核细胞。在该技术中，用抗 T 细胞抗体处理经 Percoll 分离的细胞。通过粘附到涂有亲和纯化山羊抗小鼠 IgG 的培养皿上选择性去除污染的 T 细胞。使用该步骤嗜碱性粒细胞纯度提高到34±15%。这些实验表明，阴性选择技术可使嗜碱性粒细胞纯度提高50至100倍。纯化的嗜碱性粒细胞含有预期量的组胺（1.36 - 1.54 pg/嗜碱性粒细胞），并且与未纯化的细胞一样，对不同浓度的山羊抗人 IgE Fc 产生相同量的组胺释放反应。
