Ebisawa M, Saito H, Reason D C, Sakaguchi N, Katsunuma T, Iikura Y
Division of Allergy, National Children's Medical Research Center, Tokyo, Japan.
Int Arch Allergy Appl Immunol. 1991;94(1-4):71-3. doi: 10.1159/000235328.
When cord blood mononuclear cells were cultured in the presence of rhIL-3 for 5 weeks or more, 40-90% of cultured cells became morphologically mature basophils. We analyzed the kinetics of histamine release, changes in filament actin (F-actin), and movement of intracellular Ca2+ ([Ca2+]i) induced by IgE-dependent (anti-IgE) and -independent (fMLP) stimuli in these cultured basophils. Anti-IgE and fMLP released 24.5 +/- 5.4% and 14.5 +/- 4.5% histamine from the cells, respectively. Anti-IgE caused actin polymerization with a peak response at 15 min, which began much later than the elevation of [Ca2+]i. In contrast to anti-IgE stimulation, fMLP induced rapid actin polymerization with a peak response at 30 s in correlation with kinetics of histamine release. Our results indicate that cord blood-derived cultured basophils show similar cell functions to mature basophils, and are useful models with which to investigate the mechanisms of degranulation, specifically when a large amount of highly purified cells are required.
当脐血单个核细胞在重组人白细胞介素-3(rhIL-3)存在的情况下培养5周或更长时间时,40%-90%的培养细胞在形态上会成熟为嗜碱性粒细胞。我们分析了这些培养的嗜碱性粒细胞中,由IgE依赖性(抗IgE)和非依赖性(fMLP)刺激诱导的组胺释放动力学、丝状肌动蛋白(F-肌动蛋白)的变化以及细胞内钙离子([Ca2+]i)的移动情况。抗IgE和fMLP分别从细胞中释放出24.5±5.4%和14.5±4.5%的组胺。抗IgE导致肌动蛋白聚合,在15分钟时出现峰值反应,这比[Ca2+]i的升高开始时间要晚得多。与抗IgE刺激不同,fMLP诱导快速的肌动蛋白聚合,在30秒时出现峰值反应,与组胺释放动力学相关。我们的结果表明,脐血来源的培养嗜碱性粒细胞表现出与成熟嗜碱性粒细胞相似的细胞功能,并且是用于研究脱颗粒机制的有用模型,特别是在需要大量高度纯化细胞的情况下。