Prin L, Capron M, Tonnel A B, Bletry O, Capron A
Int Arch Allergy Appl Immunol. 1983;72(4):336-46. doi: 10.1159/000234893.
Considerable variations in the distribution of eosinophil populations were revealed by density gradient centrifugation of peripheral blood leukocytes from 14 normal subjects and 31 patients with a transient or persistent hypereosinophilia. Such a purification procedure enabled us to collect in healthy controls, eosinophils (77.1 +/- 15.9% SD purity) with an appreciable cell recovery (21.7 +/- 14.0% SD) in the densest gradients ('normodense' cells). A high degree of purity was obtained in the same gradients, with leukocytes from patients with hypereosinophilia (84.4 +/- 11.9% SD pure eosinophils) but the cell recovery was significantly decreased (4.1 +/- 3.4% SD; p less than 0.001). A study of the various fractions found to contain eosinophils showed cells with an altered cellular density ('hypodense' cells) especially in marked hypereosinophilia. Furthermore, studies on leukocytes from patients with a very high hypereosinophilia (differential cell count greater than 70%) led to the recovery of almost pure fractions of eosinophils in the low density gradients. Functional studies were performed on each distinct cell fraction collected. They included investigations of their cytotoxic ability in a heterologous antibody-dependent cell-mediated cytotoxicity assay and their biochemical activity by using a previously described technique evaluating the membrane hexose transport. Variability in the functional potentialities was observed in relation to the cellular density: higher cytotoxic eosinophil abilities were noted in the case of the 'hypodense' cell population. According to biochemical criteria, these latter cells appear to be more activated and capable of responding to stimulation.
通过对14名正常受试者和31名患有短暂性或持续性嗜酸性粒细胞增多症患者的外周血白细胞进行密度梯度离心,发现嗜酸性粒细胞群体的分布存在显著差异。这样的纯化程序使我们能够在健康对照中,在最密梯度(“正常密度”细胞)中收集到纯度可观(77.1±15.9%标准差纯度)且细胞回收率可观(21.7±14.0%标准差)的嗜酸性粒细胞。在相同梯度中,嗜酸性粒细胞增多症患者的白细胞可获得高度纯度(84.4±11.9%标准差纯嗜酸性粒细胞),但细胞回收率显著降低(4.1±3.4%标准差;p<0.001)。对发现含有嗜酸性粒细胞的各个组分的研究表明,细胞密度发生改变的细胞(“低密度”细胞)尤其在明显的嗜酸性粒细胞增多症中出现。此外,对嗜酸性粒细胞增多症非常高(分类细胞计数大于70%)患者的白细胞研究导致在低密度梯度中回收了几乎纯的嗜酸性粒细胞组分。对收集的每个不同细胞组分进行了功能研究。这些研究包括在异源抗体依赖性细胞介导的细胞毒性试验中对其细胞毒性能力的研究,以及通过使用先前描述的评估膜己糖转运的技术对其生化活性的研究。观察到功能潜力与细胞密度有关的变异性:在“低密度”细胞群体中观察到更高的嗜酸性粒细胞细胞毒性能力。根据生化标准,后一种细胞似乎更具活性且能够对刺激作出反应。