Development and application of the control methods against alveolar bone resorption using human monoclonal antibody

人单克隆抗体控制牙槽骨吸收方法的开发及应用

• 批准号: 13557192
• 负责人: NISHIHARA Tatsuji
• 金额: $ 6.98万
• 依托单位: Kyushu Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557192/
• 关键词: human monoclonal antibody; periodontitis; inflammatory bone resorption; inflammatory cytokine; RANKL; osteoclast; electroporation; alveolar bone resorption; 炎症性骨吸収; 歯周病; 炎症性サイトカイン; インターロイキン-1

Osteoclasts, bone resorbing multinucleated cells, develop from monocyte-macrophage lineage cells in the presence of receptor activator NF-kB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). M-CSF-dependent bone marrow macrophages from mouse bone marrow cells have been shown to differentiate into osteoclast-like multinucleated cells (OCLs) in the presence of soluble RANKL and M-CSF. To investigate human biological responses, it is better to use human-derived proteins, such as human recombinant proteins and monoclonal antibodies. In the present study, we established human hybridomas secreting antibodies against RANKL using in vivo immunization of peripheral blood lymphocytes, and obtained some positive clones. We found that human anti-RANKL monoclonal antibody partially suppress the formation of OCL in vitro. Next, we tried to establish a method of in vivo electroporation to control the inflammatory bone resorption using human anti-RANKL antibody. However, we have no positive results in this study. Further work is needed to put a human monoclonal antibody against RANKL to practical use. Taken together, these results suggest that human monoclonal antibody against RANKL may be useful to control the inflammatory bone resorption in periodontitis.

破骨细胞是一种骨吸收多核细胞，在受体激活因子NF-κ B配体（RANKL）和巨噬细胞集落刺激因子（M-CSF）存在下由单核-巨噬细胞谱系细胞发育而来。在存在可溶性RANKL和M-CSF的情况下，来自小鼠骨髓细胞的M-CSF依赖性骨髓巨噬细胞已被证明可分化为破骨细胞样多核细胞（OCL）。为了研究人类的生物学反应，最好使用人源性蛋白质，如人重组蛋白和单克隆抗体。在本研究中，我们建立了分泌抗RANKL抗体的人杂交瘤细胞，采用体内免疫外周血淋巴细胞，并获得了一些阳性克隆。我们发现人抗RANKL单克隆抗体在体外部分抑制OCL的形成。接下来，我们尝试建立一种使用人抗RANKL抗体的体内电穿孔方法来控制炎性骨吸收。然而，我们在这项研究中没有积极的结果。需要进一步的工作将抗RANKL的人单克隆抗体投入实际应用。综上所述，这些结果表明，抗RANKL的人单克隆抗体可能有助于控制牙周炎中的炎性骨吸收。

项目成果

Nonaka, K., A.Ishisaki, N.Okahashi, T.Koseki, S.Kato, M.Muro, K.Nakashima, T.Nishihara, and Y.Kowashi: "Involvement of caspases in apoptotic cell death of macrophages infected with Actinobacillus actinomycetemcomitans"J.Periodont.Res.. 36. 40-47 (2001)

Nonaka, K.、A.Ishisaki、N.Okahashi、T.Koseki、S.Kato、M.Muro、K.Nakashima、T.Nishihara 和 Y.Kowashi：“半胱天冬酶参与巨噬细胞感染的凋亡细胞死亡

Kitamura, C., Y.Ogawa, T.Nishihara, T.Morotomi, and M.Terashita: "Transient colocalization of c-Jun N-terminal kinase and c-Jun with heat shock protein 70 in pulp cells during apoptosis"J.Dent.Res.. Accepted. (2003)

Kitamura, C.、Y.Okawa、T.Nishihara、T.Morotomi 和 M.Terashita：“细胞凋亡期间 c-Jun N 末端激酶和 c-Jun 与热休克蛋白 70 的瞬时共定位”J.

Hashimoto et al.: "Intracellular apoptosis-inducing factor is induced by a vacuolar type H+-ATPase inhibitor in B lineage cells"J Cellular Physiology. 186. 65-72 (2001)

Hashimoto 等人：“B 谱系细胞中的细胞内凋亡诱导因子是由液泡型 H-ATP 酶抑制剂诱导的”J Cellular Physiology。

Koseki, T., Y.Gao, N.Okahashi, Y.Murase, T.Tsujisawa, T.Sato, K.Yamato, and T.Nishihara: "Role of TGF-β family in osteoclastogenesis induced by RANKL"Cell Signal. 14. 31-36 (2001)

Koseki, T.、Y.Gao、N.Okahashi、Y.Murase、T.Tsujisawa、T.Sato、K.Yamato 和 T.Nishihara：“TGF-β 家族在 RANKL 诱导的破骨细胞生成中的作用”细胞信号。 14. 31-36 (2001)

Nishihara et al.: "Effect of aging on interleukin-6 release for human gingival fibroblasts stimulated with in terleukin-1B"Dentistry in Japan. 37. 20-25 (2001)

Nishihara 等人：“衰老对用白细胞介素 1B 刺激的人牙龈成纤维细胞释放白细胞介素 6 的影响”日本牙科。