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J. Biol. Chem., Vol. 278, Issue 5, 3257-3264, January 31, 2003

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CD3 Establishes a Functional Link between the T Cell Receptor and CD8^*

Marie-Agnès Doucey^§, Laurence Goffin^§¶, Dieter Naeher, Olivier Michielin^¶, Petra Baumgärtner^¶, Philippe Guillaume^¶, Ed Palmer, and Immanuel F. Luescher^¶**

From the  Institute for Biochemistry, University of Lausanne, Epalinges 1066, Switzerland, the ^¶ Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges 1066, Switzerland, and the  Laboratory of Transplantation Immunology and Nephrology, University Hospital Basel, Basel 4031, Switzerland

T cells expressing T cell receptor (TCR) complexes that lack CD3, either due to deletion of the CD3 gene, or by replacement of the connecting peptide of the TCR chain, exhibit severely impaired positive selection and TCR-mediated activation of CD8 single-positive T cells. Because the same defects have been observed in mice expressing no CD8 or tailless CD8, we examined whether CD3 serves to couple TCR·CD3 with CD8. To this end we used T cell hybridomas and transgenic mice expressing the T1 TCR, which recognizes a photoreactive derivative of the PbCS 252-260 peptide in the context of H-2K^d. We report that, in thymocytes and hybridomas expressing the T1 TCR·CD3 complex, CD8 associates with the TCR. This association was not observed on T1 hybridomas expressing only CD8 or a CD3 variant of the T1 TCR. CD3 was selectively co-immunoprecipitated with anti-CD8 antibodies, indicating an avid association of CD8 with CD3. Because CD8 is a raft constituent, due to this association a fraction of TCR·CD3 is raft-associated. Cross-linking of these TCR-CD8 adducts results in extensive TCR aggregate formation and intracellular calcium mobilization. Thus, CD3 couples TCR·CD3 with raft-associated CD8, which is required for effective activation and positive selection of CD8⁺ T cells.

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