Roles of macrophage C-type lectin in cellular trafficking of normal and malignant cells

巨噬细胞C型凝集素在正常和恶性细胞的细胞运输中的作用

• 批准号: 05671813
• 负责人: IMAI Yasuyuki
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671813/
• 关键词: Macrophages; MMGL; Lectin; Monoclonal antibody; Cancer metastasis; Trafficking; モノクロナル抗体; リンパ節

Aim of this project is to elucidate biological roles of mouse macrophage C-type lectin specific for galactose and N-acetylgalactosamine (MMGL) and to study a possible contribution of this molecule on cellular trafficking of normal and malignant cells. In order to establish topographical information on localization of MMGL-positive macrophages, we produced rat monoclonal antibodies (mAbs) specific for MMGL.We developed a novel carbohydrate binding assay based on an ELISA format to facilitate mAb screening. We systematically screened for sites of expression of MMGL using these mAbs by means of immunoblot and immunohistological analyzes, and found selective localization of MMGL-positive macrophages in connective tissue. We also detected MMGL-positive macrophages infiltrating in the metastatic nodules produced by metastatic mouse ovarian tumor cells in lung. Furthermore, we also demonstrated that MMGL is responsible for the recognition of truncated O-linked sugar chains displayd on tumor cells. In order to study possible contribution of MMGL to cellular trafficking as a cellular receptor, we transfected cDNA encoding MMGL to CTLL-2 mouse T cell line and asked whether properties of cellular trafficking could be changed. The expression of MMGL molecules was confirmed by immunoblot and flow-cytometry using mAbs. We found preferential localization of the MMGL-transfected cells within the metastatic noduled of mouse ovarian tumor cells upon injection of these cells labeled with fluorescent probes.

该项目的目的是阐明小鼠巨噬细胞 C 型凝集素对半乳糖和 N-乙酰半乳糖胺 (MMGL) 的特异性生物学作用，并研究该分子对正常和恶性细胞的细胞运输的可能贡献。为了建立 MMGL 阳性巨噬细胞定位的拓扑信息，我们生产了 MMGL 特异性的大鼠单克隆抗体 (mAb)。我们开发了一种基于 ELISA 格式的新型碳水化合物结合测定法，以促进 mAb 筛选。我们通过免疫印迹和免疫组织学分析，使用这些单克隆抗体系统地筛选了 MMGL 的表达位点，并发现 MMGL 阳性巨噬细胞在结缔组织中选择性定位。我们还检测到MMGL阳性巨噬细胞浸润在小鼠肺转移性卵巢肿瘤细胞产生的转移结节中。此外，我们还证明 MMGL 负责识别肿瘤细胞上显示的截短的 O-连接糖链。为了研究 MMGL 作为细胞受体对细胞运输的可能贡献，我们将编码 MMGL 的 cDNA 转染至 CTLL-2 小鼠 T 细胞系，并询问细胞运输的特性是否可以改变。使用 mAb 通过免疫印迹和流式细胞术证实了 MMGL 分子的表达。我们发现在注射用荧光探针标记的这些细胞后，MMGL转染的细胞优先定位在小鼠卵巢肿瘤细胞的转移结节内。

项目成果

Imai, Y.and Irimura, T.: "Quantitative measurement of carbohydrate binding activity of mouse macrophage lectin." J.Immunol.Methods. 171. 23-31 (1994)

Imai, Y. 和 Irimura, T.：“小鼠巨噬细胞凝集素碳水化合物结合活性的定量测量。”

Sakamaki, T., Imai, Y.and Irimura, T.: "Enhancement in accessibility to macrophages by modification of mucin-type carbohydrate chains on a tumor cell line : role of a C-type lectin of macrophages." J.Leukoc.Biol.57 (in press). (1995)

Sakamaki, T.、Imai, Y. 和 Irimura, T.：“通过修饰肿瘤细胞系上的粘蛋白型碳水化合物链来增强巨噬细胞的可及性：巨噬细胞 C 型凝集素的作用。”

Sakamaki,T.: "Enhancement in accessibility to macrophages by modification of mucin-type carbohydrate chains on a tummor cell line:role of a C-type lectin of macrophages." J.Leukoc.Biol.57(印刷中). (1995)

Sakamaki, T.：“通过修饰肿瘤细胞系上的粘蛋白型碳水化合物链来增强巨噬细胞的可及性：巨噬细胞 C 型凝集素的作用。”J.Leukoc.Biol.57（出版中）。 ）

Imai,Y.: "Quantitative measurement of carbohydrate binding activity of mouse macrophage lectin." J.Immunol.Methods. (印刷中).

Imai，Y.：“小鼠巨噬细胞凝集素的碳水化合物结合活性的定量测量。”J.Immunol.Methods。

Imai, Y.: "Quantitative measurement of carbohydrate binding avtivity of mouse macrophage lectin." J.Immunol. Methods. 171. 23-31 (1994)

Imai, Y.：“小鼠巨噬细胞凝集素碳水化合物结合活性的定量测量。”