用免疫荧光标记对人组织中肥大细胞亚型的分选与形态观察

方法:利用中性蛋白酶成分、特征性酶抗体的免疫荧光染色和流式细胞仪确定分选肥大细胞亚型,以激光扫描共聚焦显微镜显示肥大细胞内分泌颗粒。结果:三种免疫表型被确定:肥大细胞的类胰蛋白酶阳性(MCT)、类糜蛋白酶阴性;类糜蛋白酶阳性(MCC)、类胰蛋白酶阴性和类胰蛋白酶阳性、类糜蛋白酶阳性(MCTC)。肥大细胞内分泌颗粒分散或聚集存在,分泌颗粒突起分泌或以分散的方式释放。分泌颗粒大范围释放后,肥大细胞的形态结构发生了改变。结论:利用肥大细胞的特征性酶抗体、免疫荧光标记和流式细胞仪可将人组织中的肥大细胞分选纯化为三种亚型;以共聚焦显微镜显示肥大细胞含有丰富的分泌颗粒,它说明肥大细胞具备了为人体I型变态反应提供快速反应的物质基础。

Sorting and Morphological Observation of Mast Cell Subsets in Human Tissue with Technique of Immunofluorescence Staining

Method:Mast cell subsets were defined based on neutral protease composition and immunofluorescence staining with antibodies of specific enzyme and sorting of flow cytomery.Secretory granule in mast cell was displayed by laser scanning confocal microscopy.Results:Three immunophenotypes were determined: MCs positive for tryptase(MCT),no immunodetectable chymase;MCs positive for chymase(MCC),no immunodetectable tryptase;and MCs positive for tryptase and chymase(MCTC).There were dispersively or densely secretary vesicles in mast cell.Secretary vesicles were secreted by protuberant mode or by dispersive fashion.After large range releasing of secretary vesicles,morphological structure of mast cell would be changed.Conclusions:Three subsets of mast cell in human tissue are sorted and depurated with antibodies of specific enzyme and immunofluorescence labelling and flow cytometry.Mast cell of containing profuse secretary vacuoles could be displayed by confocal microscopy.It demonstrate that mast cell have material substructure offering for fast reaction of I type allergies of human being.