Heymanns J, Behrendt H, Schmutzler W
Agents Actions. 1982 Apr;12(1-2):192-8. doi: 10.1007/BF01965143.
Morphologically and functionally intact mast cells were isolated from the lung and mesentery of normal or actively sensitized dogs using the pronase or collagenase tissue dissociation methods. The latter method yielded about 6 times as many metachromatically staining cells. Electron microscopic examination revealed the presence of both mast cells and basophils in all samples, independent of the enzyme used for tissue dissociation. The average histamine content of the cells obtained with the pronase method was significantly higher (21.8 +/- 3.1 pg) than after collagenase treatment (16.2 +/- 4.2 pg). There was no appreciable difference in the reactivity to secretory stimulation of the cells obtained by the two methods. The cells isolated from actively sensitized dog tissues showed a significantly higher spontaneous histamine release (12.9 +/- 1.2%) than those from non-sensitized animals (7.8 +/- 1.3%) and responded equally well to challenge with both the antigens ovalbumin and horse serum. In contrast to those from normal animals, the mast cells from actively sensitized dogs released histamine on stimulation with acetylcholine, metacholine or atropine. In addition, the response to the threshold concentration of compound 48/80 (10(-6)) was significantly stronger in the sensitized cells. Small molecular polyvinylpyrrolidone (K25) was effective in mast cells from normal and actively sensitized dogs.
使用链霉蛋白酶或胶原酶组织解离方法,从正常或主动致敏犬的肺和肠系膜中分离出形态和功能完整的肥大细胞。后一种方法产生的异染性染色细胞数量约为前者的6倍。电子显微镜检查显示,所有样本中均存在肥大细胞和嗜碱性粒细胞,与用于组织解离的酶无关。用链霉蛋白酶法获得的细胞的平均组胺含量(21.8±3.1皮克)明显高于胶原酶处理后(16.2±4.2皮克)。两种方法获得的细胞对分泌刺激的反应性没有明显差异。从主动致敏犬组织中分离的细胞显示出自发性组胺释放(12.9±1.2%)明显高于未致敏动物(7.8±1.3%),并且对卵清蛋白和马血清这两种抗原的刺激反应相同。与正常动物的肥大细胞不同,主动致敏犬的肥大细胞在乙酰胆碱、醋甲胆碱或阿托品刺激下释放组胺。此外,致敏细胞对化合物48/80阈值浓度(10⁻⁶)的反应明显更强。小分子聚乙烯吡咯烷酮(K25)对正常和主动致敏犬的肥大细胞均有效。