ラット破骨細胞に対する単クローン抗体に関する研究
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概要
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The purpose of this paper is to obtain the monoclonal antibodies to rat osteoclasts. Osteoclast-rich fraction used as antigen in in vitro immunization was prepared from femora and tibia of neonatal rats. Mouse spleen cells immunized with the antigen were hybridized with myeloma cells. The culture media of the hybridoma were screened to detect the antibodies to osteoclasts. The properties of the antibodies obtained were follows : 1. In the enzyme linked immuno-staining of disaggregated bone cells, monoclonal antibodies, Cl-1 and Cl-2, reacted with osteoclasts alone. The antibody Cl-3 reacted with osteoblast-like cells and the other bone cells as well. 2. In the enzyme linked immuno-staining of bone thin section showed that Cl-1 and Cl-2 positive reactions were recognized at the resorbing part of bone surface, especially in spongy bone, and at cytoplasm of multinuclear cells nearby bone surface in bone marrow. Intense Cl-3 positive reactions appeared at all part of the bone surface and extra-cellular matrix of bone marrow. 3. Two bands, 53kDa and 47kDa, were appeared in the Western blot analysis of whole bone extract with Cl-1 or Cl-2. The antigen content, judging from the staining intesity of the bands, was high in the bone extract deprived bone marrow, and was low in bone marrow. These results show that the antibodies Cl-1 and Cl-2, likely to be a same clone, react rather specifically with osteoclasts in rat bone. The antigens reacted with Cl-1 and Cl-2 were localized in the cytoplasm of osteoclasts and the molecular weight were 53kDa and 47kDa.
- 九州歯科学会の論文
- 1995-10-25