Human skin mast cells: their dispersion, purification, and secretory characterization.
Human skin mast cells: their dispersion, purification, and secretory characterization.
复制标题
人类皮肤肥大细胞：其分散、纯化和分泌特性。
作者：
R. Benyon;M. Lowman;M. Church
用胶原酶和透明质酸酶消化人包皮，每克组织可分散约3.4×10(7)个有核细胞，其中肥大细胞占4.7%。用密度梯度离心法可将其提纯至80%。大多数肥大细胞(79%)直径在9-13微米之间，组胺含量平均为4.6pg/cell。在功能研究中，93%的细胞台盼蓝排斥和低于7%的自发性组胺分泌证明了细胞的活性。抗-IgE在5-7分钟内释放高达17.5%的细胞相关组胺。当组胺释放量为28.6%时，0.3微米的A23187对钙离子载体的释药效果最好。与人类肺肥大细胞不同，皮肤肥大细胞在化合物48/80和多聚L赖氨酸的作用下释放组胺。这种释放在20秒内完成，完全依赖于完整的糖酵解和氧化磷酸化，部分依赖于细胞外钙。P物质和吗啡诱导的分泌也有相同的特征。P物质受体的弱活性提示，P物质受体与大鼠肥大细胞一样，并不是经典的平滑肌类型。纳洛酮部分阻断了吗啡诱导的分泌，其方式与经典阿片受体的竞争性拮抗不相容。皮肤肥大细胞对非免疫刺激的敏感性明显区别于肺和淋巴组织的肥大细胞，并提供了人类肥大细胞功能异质性的证据。
Digestion of human foreskin with collagenase and hyaluronidase disperses approximately 3.4 X 10(7) nucleated cells per gram of tissue, of which mast cells constitute 4.7%. These may be purified to 80% by use of density gradient centrifugation. The majority of mast cells (79%) measured between 9 and 13 micron in diameter, and the mean histamine content was 4.6 pg/cell. Viability was demonstrated by trypan blue exclusion by 93% of the cells and the low spontaneous histamine secretion of less than 7% in functional studies. Anti-IgE released up to 17.5% of cell-associated histamine within 5 to 7 min. Calcium ionophore-induced release was optimal with 0.3 microM A23187 when 28.6% histamine was released. Unlike human lung mast cells, skin mast cells released histamine in response to compound 48/80 and poly-L-lysine. This release, which was complete within 20 sec, was totally dependent on intact glycolysis and oxidative phosphorylation and partially dependent on extracellular calcium. The same characteristics were observed with secretion induced by substance P and morphine. The weak activity of eledoisin and physalaemin suggests that the substance P receptor, like that of the rat mast cell, is not of the classical types described for smooth muscle. Morphine-induced secretion was partially blocked by naloxone in a manner not compatible with competitive antagonism at a classical opioid receptor. The sensitivity of skin mast cells to nonimmunologic stimulation clearly distinguishes them from mast cells of the lung and lymphoid tissues and provides evidence of functional heterogeneity within human mast cells.