MacGlashan D W, Schleimer R P, Peters S P, Schulman E S, Adams G K, Newball H H, Lichtenstein L M
J Clin Invest. 1982 Oct;70(4):747-51. doi: 10.1172/jci110670.
Although mediator release from mast cells and basophils plays a central role in the pathogenesis of human allergic disease, biochemical studies have been restricted to rat peritoneal mast cells and basophilic leukemia cells because they could be easily purified. We have used two new techniques of cell separation to purify human lung mast cells to 98% homogeneity. Lung cell suspensions were obtained by dispersion of chopped lung tissue with proteolytic enzymes. Mast cells were then purified from the suspensions by countercurrent centrifugal elutriation and affinity chromatography. The purified mast cells released both histamine and slow-reacting substance of anaphylaxis (SRS-A) (leukotriene C and D) during stimulation with goat anti-human IgE antibody. Moreover, these preparations were able to generate significant quantities of SRS-A (32 +/- 7 x 10(-17) LTD mole-equivalents/mast cell) at all stages of purification, indicating that a secondary cell is not necessary for the antigen-induced release of SRS.
尽管肥大细胞和嗜碱性粒细胞释放介质在人类过敏性疾病的发病机制中起着核心作用,但生化研究一直局限于大鼠腹膜肥大细胞和嗜碱性白血病细胞,因为它们易于纯化。我们使用了两种新的细胞分离技术,将人肺肥大细胞纯化至98%的同质性。通过用蛋白水解酶分散切碎的肺组织获得肺细胞悬液。然后通过逆流离心淘析和亲和层析从悬液中纯化肥大细胞。纯化的肥大细胞在山羊抗人IgE抗体刺激下释放组胺和过敏反应慢反应物质(SRS-A)(白三烯C和D)。此外,这些制剂在纯化的各个阶段都能够产生大量的SRS-A(32±7×10⁻¹⁷LTD摩尔当量/肥大细胞),这表明抗原诱导的SRS释放不需要辅助细胞。