Maeda H, Kukita T, Akamine A, Iijima T
Department of Conservative Dentistry, Faculty of Dentistry, Kyushu University, Fukuoka.
Fukuoka Igaku Zasshi. 1993 Nov;84(11):453-6.
In mammals, it has been difficult to collect the authentic osteoclasts on a large scale. Recently we established a culture system for forming osteoclast-like multinucleated cells (MNCs) which exhibited many characteristics of osteoclasts. MNCs instead of authentic osteoclasts were used as immunogen to establish the hybridomas which secrete monoclonal antibodies against the osteoclasts by in vitro immunization. We obtained two monoclonal antibodies, HOK 1 and HOK 2. HOK 1 showed intense immunoreactivity with MNCs, mononuclear cells and putative migratory traces of MNCs on the culture dishes, but it had weak reactivity with the stromal cells. HOK 2 also showed strong reactivity with MNCs, mononuclear cells, and the very limited area of the culture dishes just facing to one side of some MNCs. The stromal cells were faintly stained with HOK 2. In th paraffin sections of tibiae, both antibodies intensely stained osteoclasts, osteoblasts and osteocytes. The bone matrix was weakly stained by HOK 2 but not by HOK 1. The present observations indicated that HOK 1 and HOK 2 could recognize the common antigen expressed on cells both involving in the bone formation and resorption. The antigens recognized by HOK 1 and HOK 2 were shown to exist on the putative traces of motile MNCs. These antibodies would be available to investigate the mechanisms of the 'bone remodeling' as a functional marker of bone cells.
在哺乳动物中，大规模收集真正的破骨细胞一直很困难。最近，我们建立了一种培养系统，用于形成具有破骨细胞许多特征的破骨细胞样多核细胞（MNCs）。使用MNCs而非真正的破骨细胞作为免疫原，通过体外免疫来建立分泌抗破骨细胞单克隆抗体的杂交瘤。我们获得了两种单克隆抗体，HOK 1和HOK 2。HOK 1与MNCs、单核细胞以及培养皿上MNCs的假定迁移痕迹呈现强烈的免疫反应性，但与基质细胞的反应较弱。HOK 2也与MNCs、单核细胞以及培养皿上仅面对某些MNCs一侧的非常有限的区域呈现强烈反应性。基质细胞被HOK 2轻微染色。在胫骨的石蜡切片中，两种抗体均强烈染色破骨细胞、成骨细胞和骨细胞。骨基质被HOK 2轻微染色，但未被HOK 1染色。目前的观察结果表明，HOK 1和HOK 2可以识别参与骨形成和吸收的细胞上表达的共同抗原。HOK 1和HOK 2识别的抗原显示存在于运动性MNCs的假定痕迹上。这些抗体可作为骨细胞的功能标记物，用于研究“骨重塑”的机制。
