Identification and Functional Analysis of Cytokine Receptors on Osteoblast and Osteoclast by Monoclonal Antibodies

单克隆抗体对成骨细胞和破骨细胞细胞因子受体的鉴定及功能分析

• 批准号: 01480429
• 负责人: SUZUKI Akitoshi
• 金额: $ 4.16万
• 依托单位: Niigata University School of Dentistry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480429/
• 关键词: Cell fusion; 1alpha, 25-Dihydroxyvitamin D; Interferon-gamma; Osteoclast; Activated macrophage; F4; 80 antigen; Macrophage variant cell line; Hybridoma; 多核細胞形成; 活性型ビタミンD3; インタ-フェ-ロン-γ; モノクロ-ナル抗体; サイトカインレセプタ-

The purpose of this study is to establish the monocyte/macrophage-like cell lines which are sensitive to potent systemic and local factors, 1alpha, 25-dihydroxyvitamin D_3 (1alpha, 25 (OH)_2VD_3) and interferon-gamma (IFN-gamma). We established two variant mouse macrophage-like cell lines, whose responses to 1alpha, 25(OH)_2VD_3 and IFN-gamma differed from one another. The AH-sensitive mutant cell line (G3) was induced by allowing P388Dl tolerant to 8-azaguanine. G3 mutant cells were then fused with the 1alpha, 25(OH)_2VD_3-stimulated bone marrow cells isolated from DBA/2 mice. After AH selection the hybrid cell line (XC) was established. The G3 mutant cell line and the XC hybrid cell line had macrophage-like characteristics, such as surface antigens, Fc receptor, C3 receptor, and lysosomal enzymes. The treatment of G3 mutant cells with 1alpha, 25(OH)_2VD_3 inhibited cell proliferation with morphological changes, and increased acid phosphatase activity, phagocytic activity, and F4/80 antigen expression on the cell surface. In contrast, IFN-gamma inhibited cell proliferation without effect on acid phosphatase activity and phagocytic activity but increased F4/80 antigen expression. In XC hybrid cells, on the other hand, IFN-gamma, but not 1alpha, 25(OH)_2VD_3, inhibited cell proliferation with morphological changes but increased phagocytic activity and F4/80 antigen expression. In addition, IFN-gamma, but not 1alpha, 25(OH)_2VD_3, dose-dependently increased multinucleated cell formation of both cells. These findings suggest that the G3 mutant cell line with macrophage-like characteristics is 1alpha, 25(OH)_2VD_3-and IFN-gamma-sensitive, and that the XC hybrid cell line is, despite its macrophage-like characterisitics, only IFN-gamma-sensitive. Therefore, these newly established cell lines will provide useful systems in studying the differentiation of monocyte/macrophage lineage.

本研究的目的是建立对强大的全身和局部因子1α，25二羟基维生素D_3(1α，25(OH)_2VD_3)和干扰素-γ(干扰素-γ)敏感的单核/巨噬细胞样细胞系。我们建立了两个变异的小鼠巨噬细胞样细胞系，它们对1α、25(OH)2VD3和干扰素-γ的反应各不相同。通过P388Dl对8-氮鸟嘌呤的耐受性，获得了对8-氮鸟嘌呤敏感的突变体G3。G3突变细胞与1α，25(OH)2VD3刺激的DBA/2小鼠骨髓细胞融合。经AH筛选，建立了杂交细胞系(XC)。G3突变细胞系和XC杂交细胞系具有巨噬细胞样特性，如表面抗原、Fc受体、C3受体和溶酶体酶。1α，25(OH)2VD3处理G3突变细胞后，细胞增殖受到抑制，细胞形态发生改变，细胞表面酸性磷酸酶活性、吞噬活性和F4/80抗原表达增加。相反，干扰素-γ抑制细胞增殖，对酸性磷酸酶活性和吞噬活性无影响，但增加F4/80抗原的表达。另一方面，在XC杂交瘤细胞中，干扰素-γ(而不是1α，25(OH)_2VD3)可抑制细胞的增殖并引起细胞形态的改变，但可促进吞噬功能和F4/80抗原的表达。此外，干扰素-γ可剂量依赖性地促进两种细胞的多核细胞形成，而不是1α，25(OH)_2VD_3。这些发现表明，具有巨噬细胞样特征的G3突变细胞系对1α、25(OH)2VD3和干扰素-γ敏感，而XC杂交细胞系尽管具有巨噬细胞样特征，但只对干扰素-γ敏感。因此，这些新建立的细胞系将为研究单核/巨噬细胞系的分化提供有用的系统。

项目成果

MICHIAKI ORIKASA,TOMOYUKI KAWASE, FUJIO SHIMIZU AND AKITOSHI SUZUKI: "Establishment of Murine Macrophageーlike Mutant and Hybrid Cell Lines:Comparative Analysis of the Differentiation Induced by 1α,25ーDihydroxyvitamin D_3 and Recombinant Murine Interferonー

MICHIAKI ORIKASA、TOMOYUKI KAWASE、FUJIO SHIMIZU 和 AKITOSHI SUZUKI：“鼠类巨噬细胞样突变体和杂交细胞系的建立：1α,25-二羟基维生素 D_3 和重组鼠干扰素诱导分化的比较分析-

MICHIAKI ORIKASA,TOMOYUKI KAWASE,FUJIO SHIMIZU AND AKITOSHI SUZUKI: "Establishment of Murine Macrophageーlike Mutant and Hybrid Cell Lines:Comparative Analysis of the Differentiation Induced by 1 ,25ーDihydroxyvitamin D_3 and Recombinant Murine Interferonーγ

MICHIAKI ORIKASA、TOMOYUKI KAWASE、FUJIO SHIMIZU 和 AKITOSHI SUZUKI：“鼠类巨噬细胞样突变体和杂交细胞系的建立：1,25-二羟基维生素 D_3 和重组鼠干扰素-γ 诱导分化的比较分析

Michiaki Orikasa, Tomoyuki Kawase, Fujio Shimizu and Akitoshi Suzuki :"Establishment of murine macrophage-like mutant and hybrid cell lines : comparative analysis of the differentiation induced by 1alpha, 25-dihydroxyvitamin D_3 and recombinant murine int

Michiaki Orikasa、Tomoyuki Kawase、Fujio Shimizu 和 Akitoshi Suzuki：“鼠类巨噬细胞样突变体和杂交细胞系的建立：1α、25-二羟基维生素 D_3 和重组鼠类 int 诱导分化的比较分析

Michiaki Orikasa,: "Establishment of murine macrophage-like mutant and hybrid cell lines:Comparative analysis of the differentiation induced by 1α,25-Dihydroxyvitamin D_3 and recombinant murine interferon-γ"

Michiaki Orikasa，：“小鼠巨噬细胞样突变体和杂交细胞系的建立：1α,25-二羟基维生素 D_3 和重组小鼠干扰素-γ 诱导分化的比较分析”