Igarashi Y, Kurosawa M, Ishikawa O, Miyachi Y, Saito H, Ebisawa M, Iikura Y, Yanagida M, Uzumaki H, Nakahata T
Department of Dermatology, Gunma University School of Medicine, Maebashi, Japan.
Clin Exp Allergy. 1996 May;26(5):597-602.
The mast cell is one of the important cells in the pathogenesis of allergic disorders. However, isolating human mast cells is a laborious procedure. Recently, cultured human mast cells raised from umbilical cord blood cells have become available. It is necessary to examine whether these cells are useful in investigating the role of mast cells in human diseases.
The phenotype of mast cells depends on their anatomical sites. To examine which phenotype of mast cells these cultured mast cells most closely resemble, their ability to release was investigated.
The mast cells were raised from human umbilical cord blood cells in the presence of stem cell factor and interleukin-6. To determine the mast cell subtypes, the mast cells were immunocytochemically stained for tryptase and chymase. The cultured mast cells were then stimulated with various secretagogues, and histamine release was measured by a fluorometric technique using high-performance liquid chromatography.
The immunocytochemical staining for mast cell proteases revealed that virtually all cells contained tryptase, the definitive marker of mast cells, and that about a quarter of the cells contained chymase. Anti-IgE effectively stimulated these mast cells to release histamine in a dose-dependent, time-dependent manner. The release was completed in about 30 min. One of the non-specific stimuli, calcium ionophore A23187, also induced histamine release in a dose-dependent, time-dependent manner. In contrast, compound 48/80 and substance P failed to induce histamine release from these cells.
Cultured human mast cells resemble lung mast cells in their ability to release histamine. They will help in studying the functional properties of human mast cells and may contribute to clarifying the pathophysiology of human allergic diseases.
肥大细胞是过敏性疾病发病机制中的重要细胞之一。然而,分离人肥大细胞是一个费力的过程。最近,从脐带血细胞培养而来的人肥大细胞已可获得。有必要检查这些细胞是否有助于研究肥大细胞在人类疾病中的作用。
肥大细胞的表型取决于其解剖部位。为了检查这些培养的肥大细胞最接近哪种肥大细胞表型,对其释放能力进行了研究。
在干细胞因子和白细胞介素-6存在的情况下,从人脐带血细胞培养肥大细胞。为了确定肥大细胞亚型,对肥大细胞进行免疫细胞化学染色以检测类胰蛋白酶和糜蛋白酶。然后用各种促分泌剂刺激培养的肥大细胞,并使用高效液相色谱法通过荧光技术测量组胺释放。
肥大细胞蛋白酶的免疫细胞化学染色显示,几乎所有细胞都含有类胰蛋白酶,这是肥大细胞的决定性标志物,并且约四分之一的细胞含有糜蛋白酶。抗IgE以剂量依赖性、时间依赖性方式有效刺激这些肥大细胞释放组胺。释放过程在约30分钟内完成。非特异性刺激物之一钙离子载体A23187也以剂量依赖性、时间依赖性方式诱导组胺释放。相比之下,化合物48/80和P物质未能诱导这些细胞释放组胺。
培养的人肥大细胞在释放组胺的能力方面类似于肺肥大细胞。它们将有助于研究人肥大细胞的功能特性,并可能有助于阐明人类过敏性疾病的病理生理学。