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J Biol Chem, Vol. 273, Issue 20, 12301-12306, May 15, 1998

Physical and Functional Association between Thymic Shared Antigen-1/Stem Cell Antigen-2 and the T Cell Receptor Complex

Atsushi Kosugi, Shin-ichiroh Saitoh^¶, Satoshi Noda^¶, Kensuke Miyake, Yoshio Yamashita, Masao Kimoto, Masato Ogata^¶, and Toshiyuki Hamaoka^¶

From the  School of Allied Health Sciences, Faculty of Medicine, Osaka University, Osaka 565, ^¶ Biomedical Research Center, Osaka University Medical School, Osaka 565, and  Department of Immunology, Saga Medical School, Saga 849, Japan

Thymic shared antigen-1 (TSA-1)/stem cell Ag-2 (Sca-2) is a glycosylphosphatidylinositol (GPI)-anchored antigen expressed on lymphocytes. We have previously demonstrated that a signal via TSA-1/Sca-2 inhibits T cell receptor (TCR)-mediated 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 CD3 chains in T cell hybridomas, activated T cells, and COS-7 cells transfected with TSA-1 and CD3 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 CD3 did not require the GPI anchor of TSA-1. Finally, tyrosine phosphorylation of CD3 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 CD3 underlies a regulatory role of TSA-1/Sca-2 in the TCR-signaling pathway.

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