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CD45 ectodomain controls interaction with GEMs and Lck activity for optimal TCR signaling

Nature Immunology volume 4, pages 189–197 (2003)Cite this article

Abstract

The transmembrane phosphatase CD45 regulates both Lck activity and T cell receptor (TCR) signaling. Here we have tested whether the large ectodomain of CD45 has a role in this regulation. A CD45 chimera containing the large ectodomain of CD43 efficiently rescues TCR signaling in CD45-null T cells, whereas CD45 chimeras containing small ectodomains from other phosphatases do not. Both basal Lck activity in unstimulated cells and the TCR-induced increase in tyrosine phosphorylation of the TCR ζ-chain and in Lck activity depend on the expression of CD45 with a large ectodomain. Unlike CD45 chimeras containing small ectodomains, both the CD45 chimera with a large ectodomain and wild-type CD45 itself are partially localized to glycosphingolipid-enriched membranes (GEMs). Taken together, these data show that the large CD45 ectodomain is required for optimal TCR signaling.

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Figure 1: Construction of the CD45 chimeras.
Figure 2: Reconstitution of TCR-induced NFAT activity by CD45WT.
Figure 3: Inefficient reconstitution of TCR-induced signaling by Thy-1–CD45 and CD2-CD45.
Figure 4: CD43 and CD45RO ectodomains reconstitute TCR-induced activation.
Figure 5: TCR-induced NFAT activation in stable transfectants expressing CD43-CD45, Thy-1–CD45 and CD2-CD45 chimeras.
Figure 6: Inefficient TCRζ phosphorylation and pTyr394–Lck formation in cells stably reconstituted with Thy-1–CD45 and CD2-CD45.
Figure 7: Thy-1–CD45 with a Glu613Arg mutation does not reconstitute TCR-induced NFAT activation.
Figure 8: Differential distribution of CD45 chimeras in GEM domains.

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Acknowledgements

We thank B. Schraven, A. Shaw, D. Alexander and B. Alarcon for reagents and cell lines; and G. Langsley, S. Pellegrini and H.-T. He for suggestions. This work was supported by grants from the Pasteur Institute, the Medical Research Council, the Association pour la Recherche sur le Cancer and the Conseil National de la Recherche Scientifique. C. I. was a recipient of a fellowship from the Consejo Nacional para la Ciencia y Tecnología, from Mexico.

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  1. Department of Immunology, Molecular Immunology Unit, Institut Pasteur, 25 rue du Dr. Roux, Paris, 75724, Cedex 15, France

    Claudine Irles, Frédérique Michel & Oreste Acuto

  2. Sir William Dunn School of Pathology, South Parks Road, Oxford University, Oxford, OX1 3RE, UK

    Antony Symons, Talitha R. Bakker & P. Anton van der Merwe

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Irles, C., Symons, A., Michel, F. et al. CD45 ectodomain controls interaction with GEMs and Lck activity for optimal TCR signaling. Nat Immunol 4, 189–197 (2003). https://doi.org/10.1038/ni877

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