Role of sugar chain in the induction of Fc receptor-mediated phagocytosis by macrophages

糖链在诱导巨噬细胞 Fc 受体介导的吞噬作用中的作用

• 批准号: 62580109
• 负责人: TAKASAKI Seiichi
• 金额: $ 1.22万
• 依托单位: the 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: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62580109/
• 关键词: Fc receptor-mediated phagocytosis; High mannose-type sugar chain; プロセシング阻害剤; 貧食能誘導; Fcレセプター; マクロファージ系細胞; アスパラギン結合型糖鎖; プロセシング

Phagocytosis can be separated into two steps, attachment of the foreign material to the cell membrane and ingestion of the material. A mouse macrophage-like cell line, P388Dl, expresses receptors which recognize Fc portion of IgG bound to antigen, but can not take up the antigen-antibody conplex. However, addition of dimethyl sulfoxide (DMSO) toe the culture medium induces the ability of ingestion in more than 90% of the cells after 2 days. It is known that structural changes are accompanied with macrophage differentiation. Therefore, sugar chain structures were compared before and after DMSO induction of P388DL cells, and the relationship between structural changes and induction of Fc receptor-mediated phagocytosis was studied in this project. Finally, the following results were obtained. (1) From the analysis of NP-40 extracts of the cells by SDS-PAGE, it was obvious that the total protein pattern does not change after DMSO induction. But, increased staining by Concanavalin A of glyc … More oproteins with molecular sizes of more than 30 KDa was observed after induction. (2) After DMSO induction, high mannose-type sugar chains increased with the decrease of tri- and tetraantennary complex-type sugar chains. Thus, the data suggestd that inhibition of the processing of ASN-linked sugar chains is involved in the structural change caused by DMSO induction. (3) When the cells were cultured in the presence of processing inhibitors (swainsonine and castanospermine), which accumulate high mannose-type sugar chains, Fc receptor-mediated phagocytosis was induced depending on the dose of inhibitors and the culture time.These results indicate that structural change of sugar chains is not merely accompanied with the induction, but causes the induction of Fc receptor-mediated phagocytosis. Expression of high mannose-type sguar chains in some specific glycoproteins might be a key for the induction, suggesting that it is important to analyze the mechanism of induction on the molecular basis in the near future. Less

吞噬作用可以分为两个步骤，将异物附着在细胞膜上和摄入该物质。小鼠巨噬细胞样细胞系P388 D1表达识别与抗原结合的IgG的Fc部分的受体，但不能摄取抗原-抗体复合物。然而，向培养基中添加二甲基亚砜（DMSO）在2天后诱导超过90%的细胞的摄取能力。已知结构变化伴随巨噬细胞分化。因此，本项目比较了DMSO诱导P388 DL细胞前后的糖链结构，并研究了结构变化与Fc受体介导的吞噬作用诱导之间的关系。最后，获得了以下结果。(1)通过SDS-PAGE分析细胞的NP-40提取物，很明显，DMSO诱导后总蛋白质模式没有改变。但是，随着糖基化程度的增加， ...更多信息 诱导后观察到分子大小大于30 KDa的α蛋白。(2)DMSO诱导后，高甘露糖型糖链增加，三触角和四触角复合型糖链减少。因此，数据表明，ASN连接的糖链的加工的抑制涉及由DMSO诱导引起的结构变化。(3)当细胞在含有高甘露糖型糖链的加工抑制剂（苦马豆素和栗精胺）的培养液中培养时，Fc受体介导的吞噬作用被诱导，并随抑制剂的剂量和培养时间而变化，表明糖链结构的改变不仅伴随着诱导，而且引起Fc受体介导的吞噬作用的诱导。在某些特定糖蛋白中高甘露糖型糖链的表达可能是诱导的关键，这表明在不久的将来在分子基础上分析诱导机制是重要的。少

项目成果

Keiko Fukushima: "Induction of Fc receptor-mediated phagocytosis of a mouse macrophage-like cell line P388D1 by swainsonine and castanospermine."

Keiko Fukushima：“苦豆蔻碱和粟精胺诱导小鼠巨噬细胞样细胞系 P388D1 的 Fc 受体介导的吞噬作用。”

Seiichi Takasaki: "Structural changes in sugar chains of membrane glycoproteins during hematopoietic cell differentiation." Seikagaku. 59. 205-222 (1987)

Seiichi Takasaki：“造血细胞分化过程中膜糖蛋白糖链的结构变化。”

高崎誠一: 生化学. 59. 205-222 (1987)

高崎精一：生物化学。59。205-222（1987）