Analysis of TSA-1 that regulates TCR-mediated signal transduction

调节 TCR 介导的信号转导的 TSA-1 分析

• 批准号: 08839014
• 负责人: KOSUGI Atsushi
• 金额: $ 1.22万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08839014/
• 关键词: T cells; T cell Receptor; TSA-1; src-family kinases; DIG; CD3zeta; LCK; CD35鎖; GPIアンカー型タン白質; シグナル伝達; 物理的会合

Thymic shared antigen-1 (TSA-1)/stem cell Ag-2 (Sca-2) is a glycosyl-phosphatidylinositol (GPI) -anchored antigen expressed on lymphocytes. We have previously demonstrated that a signal via TSA-1/Sca-2 inhibits T cell receptor (TCR)-medialted T cell activation and apoptosis. To elucidate a molecular mechanism for TSA-1-mediated modulation of the TCR signaling pathway, we examined whether TSA-1 is physically coupled to the TCR in the present study. TSA-1 was clearly associated with CD3zeta chains in T cell hybridomas, activated T cells, and COS-7 cells transfected with TSA-1 and CD3zeta cDNA.The physical association was confirmed on the surface of T cells in immunoprecipitation and confocal microscopy. The analysis using stable and transient transfectants expressing a transmembrane form of TSA-1 revealed that the association of CD3zeta did not require the GPI anchor of TSA-1. Finally, tyrosine phosphorylation of CD3zeta chains was induced after stimulation with anti-TSA-1, suggesting that a functional association between these two molecules also exists. These results imply that the physical association to CD3zeta underlies a regulatory role of TSA-1/Sca-2 in TCR signaling pathway.

胸腺共享抗原-1(TSA-1)/干细胞抗原-2(SCA-2)是一种表达于淋巴细胞上的糖基磷脂酰肌醇(GPI)锚定抗原。我们先前已经证明，通过TSA-1/SCA-2的信号抑制T细胞受体(TCR)介导的T细胞激活和凋亡。为了阐明TSA-1介导的TCR信号通路的分子机制，我们在本研究中研究了TSA-1是否与TCR物理偶联。在T细胞杂交瘤、活化T细胞和转染TSA-1和CD3zeta cDNA的COS-7细胞中，TSA-1明显与CD3zeta链结合，免疫沉淀和共聚焦显微镜观察证实TSA-1与CD3zeta链在T细胞表面发生物理结合。用稳定和瞬时表达TSA-1跨膜形式的转染体进行的分析表明，CD3zeta的结合不需要TSA-1的GPI锚点。最后，在抗TSA-1的刺激下，CD3zeta链的酪氨酸磷酸化被诱导，这表明这两个分子之间也存在功能联系。这些结果表明，TSA-1/SCA-2与CD3zeta的物理结合可能是TCR信号通路中的一种调节作用。

项目成果

A.Kosugi et al.: "Subunit composition of the pre-T-cell receptor complex analyzod by monoclonal antibody against the pre-T-cell receptor chain." Immunology. 91. 618-622 (1997)

A.Kosugi 等人：“通过针对前 T 细胞受体链的单克隆抗体分析前 T 细胞受体复合物的亚基组成。”

Kosugi, A., Saitoh, S.-I., Noda, S., Miyake, K., Yamashita, Y., Kimoto, M., Ogata, M., and Hamaoka, T.: "Physical and functional association between thymic shared antigen-1 (TSA-1)/stem cell antigen-2 (Sca-2) and the T cell receptor (TCR) complex." J.Biol

Kosugi, A.、Saitoh, S.-I.、Noda, S.、Miyake, K.、Yamashita, Y.、Kimoto, M.、Ogata, M. 和 Hamaoka, T.：“之间的物理和功能关联

Kosugi, A., et al.: "Physical and functional association between thymic shared antigen-I(TSA-1)lstem cell antigen -2(Sca-2)and the T cell receptor(TCR)complex." J.Biol.Chem.(印刷中). (1998)

Kosugi, A. 等人：“胸腺共享抗原 -I(TSA-1)l 干细胞抗原 -2(Sca-2) 和 T 细胞受体 (TCR) 复合物之间的物理和功能关联。”化学（出版中）。

T.Furuyama et al.: "Identification of a novel transmembrane semaphorin expressed on lymphocytes." J.Biol.Chem.271. 33376-33381 (1996)

T.Furuyama 等人：“鉴定淋巴细胞上表达的新型跨膜信号蛋白。”

Kosugi,A., et.al.: "Subunit composition of the pre-T-cell receptor complex analyzed by monoclonal antibody against the pre-T-cell receptor α chain." Immunology. 91. 618-622 (1997)

Kosugi, A. 等人：“通过针对前 T 细胞受体 α 链的单克隆抗体分析前 T 细胞受体复合物的亚基组成。免疫学”。