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The proline-rich sequence of CD3ε controls T cell antigen receptor expression on and signaling potency in preselection CD4+CD8+ thymocytes

Nature Immunology volume 9, pages 522–532 (2008)Cite this article

Abstract

Antigen recognition by T cell antigen receptors (TCRs) is thought to 'unmask' a proline-rich sequence (PRS) present in the CD3ε cytosolic segment, which allows it to trigger T cell activation. Using 'knock-in' mice with deletion of the PRS, we demonstrate here that elimination of the CD3ε PRS had no effect on mature T cell responsiveness. In contrast, in preselection CD4+CD8+ thymocytes, the CD3ε PRS acted together with the adaptor protein SLAP to promote CD3ζ degradation, thereby contributing to downregulation of TCR expression on the cell surface. In addition, analysis of CD4+CD8+ thymocytes of TCR-transgenic mice showed that the CD3ε PRS enhanced TCR sensitivity to weak ligands. Our results identify previously unknown functions for the evolutionarily conserved CD3ε PRS at the CD4+CD8+ developmental stage and suggest a rather limited function in mature T cells.

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Figure 1: Higher TCR expression on DP thymocytes from Cd3eΔPRS/ΔPRS mice.
Figure 2: The Cd3eΔPRS mutation abrogates Nck docking.
Figure 3: TCR internalization and recycling in Cd3eΔPRS/ΔPRS DP thymocytes.
Figure 4: The MHC class II–CD4–Lck–SLAP pathway and CD3ε PRS act together to downregulate αβ TCR expression on DP cells.
Figure 5: The CD3ε PRS and SLAP regulate CD3ζ degradation in DP thymocytes.
Figure 6: Impeded positive selection in Cd3eΔPRS/ΔPRS mice expressing the HY transgenic TCR.
Figure 7: The CD3ε PRS enhances the responsiveness of DP thymocytes to weak TCR stimuli.
Figure 8: Tyrosine phosphorylation in thymocytes from wild-type, Cd3eΔPRS/ΔPRS and Sla−/− mice after TCR cross-linking.

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Acknowledgements

We thank H. Boukarabila, C. Boyer, Y. Wang, S. Yamasaki, A. Tafuri, I. Prinz, H.T. He and L. Poulin for discussions; P. Grenot, M. Barad, N. Brun, M. Richelme, P. Perrin, F. Danjan and A. Gillet for advice and technical assistance; D. Gil (Mayo Clinic Rochester) for GST and GST–Nck-SH3.1 constructs; S. Yamasaki (Riken Research Center for Allergy and Immunology) for Tst4-DL1 cells; and A.M. Schmitt-Verhulst (Centre d'Immunologie de Marseille-Luminy) for biotin-conjugated anti-H-2Kb. Supported by the Centre National de la Recherche Scientifique, Institut National de la Santé et de la Recherche Médicale, European Communities (MUGEN Network of Excellence), Agence Nationale pour la Recherche (Plate-forme Technologique du Vivant IBISA-MNG), Association pour la Recherche sur le Cancer, Fondation pour la Recherche Médicale, Ecole Normale Supérieure (M.Mi.) and European Molecular Biology Organization (R.R.).

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  1. Enrique Aguado

    Present address: Present address: Hospital Universitario de Puerto Real, Unidad de Investigación, Carretera Nacional IV Km. 665, Cádiz, Spain.,

Authors and Affiliations

  1. Centre d'Immunologie de Marseille-Luminy, Université de la Méditerrannée, Case 906, Institut National de la Santé et de la Recherche Médicale U631, and Centre National de la Recherche Scientifique UMR6102, Marseille, 13288, Cedex 9, France

    Michaël Mingueneau, Amandine Sansoni, Claude Grégoire, Romain Roncagalli, Enrique Aguado, Marie Malissen & Bernard Malissen

  2. Department of Medicine, Rosalind Russell Medical Research Center for Arthritis, Howard Hughes Medical Institute, University of California San Francisco, San Francisco, 94143, California, USA

    Arthur Weiss

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Contributions

All authors contributed to discussions of experimental design and data analysis; M.Mi. did all experimental studies unless otherwise indicated; A.S. provided technical assistance; C.G. and R.R. helped with biochemistry and calcium imaging; E.A. helped with the 'knock-in' strategy; A.W. provided suggestions and mice deficient in SLAP and Zap70; M.Ma. and B.M. directed the study; and M.Mi. and B.M. wrote the manuscript.

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Correspondence to Bernard Malissen.

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Mingueneau, M., Sansoni, A., Grégoire, C. et al. The proline-rich sequence of CD3ε controls T cell antigen receptor expression on and signaling potency in preselection CD4+CD8+ thymocytes. Nat Immunol 9, 522–532 (2008). https://doi.org/10.1038/ni.1608

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