Razin E, Mencia-Huerta J M, Stevens R L, Lewis R A, Liu F T, Corey E, Austen K F
J Exp Med. 1983 Jan 1;157(1):189-201. doi: 10.1084/jem.157.1.189.
Mouse bone marrow-derived mast cells differentiated in vitro and sensitized with monoclonal IgE respond to antigen-initiated activation with the release of histamine, beta-hexosaminidase, chondroitin sulfate E proteoglycan, and leukotriene C4 (LTC4). The chondroitin sulfate E nature of the glycosaminoglycan side chain was established by demonstrating that the chondroitinase ABC disaccharide digestion products were composed of equal quantities of 4-sulfated and 4,6-disulfated N-acetyl-galactosamine. The single immunoreactive sulfidopeptide leukotriene, released and quantitated with a class-specific antibody, was identified as LTC4 by its retention time on reverse-phase high-performance liquid chromatography and by its specific spasmogenic activity on the guinea pig ileum. The release of the preformed mediators, as well as of LTC4, was related in a dose-response fashion to the concentration of monoclonal IgE used during the sensitization step and to the concentration of specific antigen used to initiate the activation-secretion response. The optimal concentrations of IgE for sensitization and of antigen for challenge were the same for the release of preformed mediators and of LTC4. In addition, the time courses of their release were superimposable, with a plateau at 5 min after antigen challenge. The release of three preformed mediators and of LTC4 after fixation of IgE, washing of the sensitized cells, and antigen challenge unequivocally indicates a bone marrow-derived mast cell origin for these products. Linear regression analyses of the net percent release of beta-hexosaminidase to histamine and of 35S-chondroitin sulfate E to beta-hexosaminidase yielded straight lines that intersected at the origin, which indicates that the three preformed mediators are localized in the secretory granules of the bone marrow-derived mast cells. The concomitant generation of 23 ng of LTC4/10(6) sensitized bone marrow-derived mast cells represents the first example of IgE-dependent release of substantial amounts of LTC4, a component of slow reacting substance of anaphylaxis, from a mast cell population of greater than 95% purity. The IgE-dependent generation of LTC4, rather than prostaglandin D2, by the chondroitin sulfate E proteoglycan-containing bone marrow-derived mast cells contrasts with the predominant generation of prostaglandin D2 by heparin proteoglycan-containing mast cells. These differences together support the existence of two phenotypically different mast cell subclasses.
体外分化并用单克隆IgE致敏的小鼠骨髓源性肥大细胞，在抗原引发激活后会释放组胺、β-己糖胺酶、硫酸软骨素E蛋白聚糖和白三烯C4（LTC4）。通过证明硫酸软骨素酶ABC二糖消化产物由等量的4-硫酸化和4,6-二硫酸化的N-乙酰半乳糖胺组成，确定了糖胺聚糖侧链的硫酸软骨素E性质。用类特异性抗体释放并定量的单一免疫反应性硫肽白三烯，通过其在反相高效液相色谱上的保留时间以及对豚鼠回肠的特异性致痉活性，被鉴定为LTC4。预先形成的介质以及LTC4的释放，与致敏步骤中使用的单克隆IgE浓度以及用于引发激活-分泌反应的特异性抗原浓度呈剂量反应关系。致敏的最佳IgE浓度和激发的最佳抗原浓度，对于预先形成的介质和LTC4的释放是相同的。此外，它们释放的时间进程是重叠的，抗原激发后5分钟达到平台期。在固定IgE、洗涤致敏细胞并进行抗原激发后，三种预先形成的介质和LTC4的释放明确表明这些产物起源于骨髓源性肥大细胞。β-己糖胺酶对组胺的净释放百分比以及35S-硫酸软骨素E对β-己糖胺酶的线性回归分析产生了在原点相交的直线，这表明三种预先形成的介质定位于骨髓源性肥大细胞的分泌颗粒中。每10(6)个致敏的骨髓源性肥大细胞伴随产生23 ng的LTC4，这代表了从纯度大于95%的肥大细胞群体中IgE依赖性释放大量LTC4（过敏反应慢反应物质的一种成分）的首个例子。含硫酸软骨素E蛋白聚糖的骨髓源性肥大细胞IgE依赖性产生LTC4而非前列腺素D2，这与含肝素蛋白聚糖的肥大细胞主要产生前列腺素D2形成对比。这些差异共同支持了两种表型不同的肥大细胞亚类的存在。
