Maeda H, Kukita T, Akamine A, Kukita A, Iijima T
Department of Periodontics and Endodontics, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
Histochemistry. 1994 Oct;102(4):247-54. doi: 10.1007/BF00269160.
The characteristics of a monoclonal antibody produced against osteoclast-like multinucleated cells (MNCs) formed in rat bone marrow cultures were examined immunohistochemically and biochemically. The in vitro immunization was performed using as immunogen the MNCs from rat bone marrow cell culture, which revealed many characteristics of osteoclasts. After screening and cloning of hybridomas, the monoclonal antibody HOK 1 was obtained. This antibody reacted weakly with stromal cells and intensely with both MNCs and their putative migratory traces on culture dishes. Immunofluorescent examination of paraffin sections revealed intense reactivity on the epithelium of the choroid plexus, the ileum and the proximal-convoluted tubules of the kidney, and also on bone cells such as osteocytes, osteoblasts, and osteoclasts. Western blotting using purified rat osteopontin verified that the antigen recognized by HOK 1 was osteopontin. Positive HOK 1 immunoreactivity was further observed in the resorption lacunae formed by a culture of MNCs on human tooth slices and on the surface of osteoclasts. The present data suggested that osteopontin is preferentially present on the resorption lacunae in resorbing calcified matrices and that osteoclasts under a specific state might trap this protein on their cell surface.
对针对在大鼠骨髓培养物中形成的破骨细胞样多核细胞（MNCs）产生的单克隆抗体的特性进行了免疫组织化学和生物化学检测。使用大鼠骨髓细胞培养物中的MNCs作为免疫原进行体外免疫，这些MNCs显示出破骨细胞的许多特征。在对杂交瘤进行筛选和克隆后，获得了单克隆抗体HOK 1。该抗体与基质细胞反应较弱，与MNCs及其在培养皿上的假定迁移痕迹反应强烈。石蜡切片的免疫荧光检查显示，脉络丛上皮、回肠和肾近端曲管以及骨细胞如骨细胞、成骨细胞和破骨细胞有强烈反应。使用纯化的大鼠骨桥蛋白进行的蛋白质印迹证实，HOK 1识别的抗原是骨桥蛋白。在人牙切片上由MNCs培养形成的吸收陷窝以及破骨细胞表面进一步观察到HOK 1阳性免疫反应性。目前的数据表明，骨桥蛋白优先存在于吸收钙化基质中的吸收陷窝上，并且处于特定状态的破骨细胞可能会将这种蛋白质捕获在其细胞表面。
