Gibson S, Miller H R
Immunology. 1986 May;58(1):101-4.
The specificities of antibodies raised in rabbits against rat mast cell proteinase 1 (RMCP 1) from connective tissue mast cells (CTMC), and against RMCP II from mucosal mast cells (MMC), were analysed by SDS-PAGE and Western blotting. Significant cross-reactivity was detected, and was eliminated by affinity purification and cross-absorption techniques. The resultant F(ab')2 antibodies, monospecific for each enzyme, were used for the immunohistochemical localization of RMCP I and II in rat tissues. Cells in skin, tongue, intestinal serosa and lung parenchyma which, by histochemical techniques, have been identified as CTMC, contained RMCP I exclusively. Cells in jejunal lamina propria and bronchial epithelium, previously classified as MMC, contained RMCP II. The results demonstrate the feasibility of distinguishing mast cell subsets by their content of serine proteinases.
通过SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹法(Western blotting)分析了兔体内产生的针对来自结缔组织肥大细胞(CTMC)的大鼠肥大细胞蛋白酶1(RMCP 1)以及针对来自黏膜肥大细胞(MMC)的RMCP II的抗体的特异性。检测到显著的交叉反应性,并通过亲和纯化和交叉吸收技术将其消除。所得的对每种酶具有单特异性的F(ab')2抗体用于RMCP I和II在大鼠组织中的免疫组织化学定位。通过组织化学技术已鉴定为CTMC的皮肤、舌头、肠浆膜和肺实质中的细胞仅含有RMCP I。空肠固有层和支气管上皮中先前分类为MMC的细胞含有RMCP II。结果证明了通过肥大细胞的丝氨酸蛋白酶含量区分肥大细胞亚群的可行性。