Establishment and Analysis of mice disrupted with DNAM-1 gene

DNAM-1基因破坏小鼠的建立及分析

• 批准号: 10833002
• 负责人: SHIBUYA Akira
• 金额: $ 1.86万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10833002/
• 关键词: DNAM-1; Adhesion Molecules; Lymphocyte; 細胞障害性リンパ球

DNAM-1 is a signal transducing adhesion molecule involved in the cytolytic function mediated by CTL and NK cells. Cross-linking DNAM-1 with anti-DNAM-1 mAb induces cytolysis mediated by CTL and NK cells and also results in tyrosine phosphorylation of the DNAM-1 molecule. We studied regulation of DNAM-1-mediated signaling and adhesion. We report here that specific inhibitors of PKC activity prevented DNAM-1-mediated cytolytic activation of NK cells. Adhesion of DNAM-1 to its ligand does not require divalent cations, such as magnesium or calcium, and is regulated by PKC, as demonstrated by augmentation of DNAM-1 adhesion by PMA and inhibition by specific PKC inhibitors. Mutation of the putative PKC binding site in the cytoplasmic domain of DNAM-1 (SerィイD1329ィエD1 to PheィイD1329ィエD1) prevents ligand binding and PMA-induced serine phosphorylation of the DNAM-1 receptor. These results indicate that PKC phosphorylates SerィイD1329ィエD1 of DNAM-1 and plays a critical role for both DNAM-1 adhesion and signaling.Whereas ligation of DNAM-1 adhesion molecule triggers cytotoxicity mediated by normal NK and T cells, this function was defective in NK cell clones from leukocyte adhesion deficiency syndrome. However, genetic reconstitution of cell surface expression of LFA-1 restored the ability of DNAM-1 to initiate anti-DNAM-1 mAb induced cytotoxicity, indicating a functional relationship between DNAM-1 and LFA-1. Further studies demonstrated that LFA-1 physically associates with DNAM-1 in NK cells and anti-CD3 mAb stimulated T cells, for which serine phosphorylation of DNAM-1 plays a critical role. In addition, cross-linking of LFA-1 induces tyrosine phosphorylation of DNAM-1, for which the fyn protein tyrosine kinase is responsible. These results indicate that DNAM-1 is involved in the LFA-1-mediated intracellular signals.

DNAM-1是由CTL和NK细胞介导的胞质函数中的信号转导分子，可诱导由CTL和NK细胞介导的胞溶性，还导致您研究了DNAM-1分子的酪氨酸磷酸化。 NAM-1介导的NK细胞粘附的介导的信号传导和粘附。通过IFIC PKC抑制剂。粘附和信号传导。DNAM-1粘附分子的连接触发由NK和TE LLS介导的细胞毒素，THOS功能在白细胞粘附综合征的NK细胞克隆中有缺陷-1 MAB诱导的细胞毒性，表明DNAM-1和LFA-1之间的功能关系表明，LFA-1在NK细胞中与DNAM-1物理相关联和抗CD3 MAB刺激的T细胞，DNAM的丝氨酸磷酸化-1起关键作用，DNAM-1的LFA-1 CES酪氨酸磷酸化的交联，因此，蛋白激酶是造成的。

项目成果

Shibuya, A, et al: "Protein kinase C is involved in the regulation of both signaling and adresion mediated DNAM-1"J. Immunology. 161. 1671-1676 (1998)

Shibuya, A, 等人：“蛋白激酶 C 参与信号传导和邻接介导的 DNAM-1 的调节”J.

Shibuya K., Lanier LL., Phillips JH., Ochs D., Shimizu K., Nakayama E., Nakauchi H., Shibuya A.: "Physical and functional association of LFA-1 with DNAM-1 adhesion molecule."Immunity. 11. 615-623 (1999)

Shibuya K.、Lanier LL.、Phillips JH.、Ochs D.、Shimizu K.、Nakayama E.、Nakauchi H.、Shibuya A.：“LFA-1 与 DNAM-1 粘附分子的物理和功能关联。”

Shibuya A., Lanier LL., Phillips JH.: "Protein kinase C is involved in the regulation of both signaling and adhesion mediated by DNAM-1 receptor."J. Immunol.. 161. 1671-1676 (1998)

Shibuya A.、Lanier LL.、Phillips JH.：“蛋白激酶 C 参与 DNAM-1 受体介导的信号传导和粘附的调节。”J.

Shibuya K.et al.: "Physical and functional association of LFA-1 with DNAM-1 adhesion molecule"Immunity. 11. 615-623 (1999)

Shibuya K.等人：“LFA-1 与 DNAM-1 粘附分子的物理和功能关联”免疫。

Shibuya,A,et al: "Protein kinase C is involved in the regulation of both signaling and adhesion mediated by DNAM-1" J.Immunology. 161. 1671-1676 (1998)

Shibuya,A,et al：“蛋白激酶 C 参与 DNAM-1 介导的信号传导和粘附的调节”J.Immunology。