Razin E, Mencia-Huerta J M, Lewis R A, Corey E J, Austen K F
Proc Natl Acad Sci U S A. 1982 Aug;79(15):4665-7. doi: 10.1073/pnas.79.15.4665.
Bone marrow-derived mast cells, differentiated in vitro, demonstrate surface IgE, receptors and contain histamine in metachromatic granules, which are composed of chondroitin sulfate E proteoglycan rather than heparin proteoglycan. Activation of this subclass of mast cells with calcium ionophore A23187 resulted in generation of immunoreactive C-6-sulfidopeptide leukotriene in a dose- and time-dependent fashion. Isolation of immunoreactive C-6-sulfldopeptide leukotriene by reverse-phase high-performance liquid chromatography (RP-HPLC) revealed a retention time and a specific biologic activity identical to those of synthetic leukotriene C4 (LTC4). Neither radiolabeled nor immunoreactive conversion products of [3H]LTC4/LTC4 were recognized during RP-HPLC resolution of the supernatants. The failure of fresh bone marrow cultures to generate C-6-sulfidopeptide leukotriene in response to ionophore indicates that leukotriene generation is dependent upon cellular differentiation into a mast cell population. The amount of LTC4 generated during optimal ionophore stimulation, 90.9 +/- 7.5 ng per 10(6) cells, contrasts with the relatively low amounts of C-6-sulfidopeptide leukotriene generated by the conventional heparin-containing rat mast cells or mouse mastocytoma cells.
体外分化的骨髓源性肥大细胞表现出表面IgE受体,并且在异染颗粒中含有组胺,这些颗粒由硫酸软骨素E蛋白聚糖而非肝素蛋白聚糖组成。用钙离子载体A23187激活这类肥大细胞会以剂量和时间依赖的方式产生免疫反应性C-6-硫代肽白三烯。通过反相高效液相色谱(RP-HPLC)分离免疫反应性C-6-硫代肽白三烯,其保留时间和特定生物学活性与合成白三烯C4(LTC4)相同。在上清液的RP-HPLC分离过程中,未识别到[3H]LTC4/LTC4的放射性标记或免疫反应性转化产物。新鲜骨髓培养物对离子载体无反应而无法产生C-6-硫代肽白三烯,这表明白三烯的产生依赖于细胞分化为肥大细胞群体。在最佳离子载体刺激下产生的LTC4量为每10(6)个细胞90.9±7.5 ng,这与传统含肝素的大鼠肥大细胞或小鼠肥大细胞瘤细胞产生的相对少量的C-6-硫代肽白三烯形成对比。