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The lymphoid lineage–specific actin-uncapping protein Rltpr is essential for costimulation via CD28 and the development of regulatory T cells

Nature Immunology volume 14, pages 858–866 (2013)Cite this article

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Abstract

Although T cell activation can result from signaling via T cell antigen receptor (TCR) alone, physiological T cell responses require costimulation via the coreceptor CD28. Through the use of an N-ethyl-N-nitrosourea–mutagenesis screen, we identified a mutation in Rltpr. We found that Rltpr was a lymphoid cell–specific, actin-uncapping protein essential for costimulation via CD28 and the development of regulatory T cells. Engagement of TCR-CD28 at the immunological synapse resulted in the colocalization of CD28 with both wild-type and mutant Rltpr proteins. However, the connection between CD28 and protein kinase C-θ and Carma1, two key effectors of CD28 costimulation, was abrogated in T cells expressing mutant Rltpr, and CD28 costimulation did not occur in those cells. Our findings provide a more complete model of CD28 costimulation in which Rltpr has a key role.

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Figure 1: Identification of Basilic Lat(Y136F) mice.
Figure 2: Characterization of the Basilic mutation and expression of Rltpr.
Figure 3: Development of conventional T cells in RltprBas and Cd28−/− mice.
Figure 4: Similar T cell functional defects in RltprBas and Cd28−/− mice.
Figure 5: Development of Treg cells in RltprBas and Cd28−/− mice.
Figure 6: Microclusters of Rltpr or RltprBas form in a CD80-dependent manner and localize together with CD28 microclusters.
Figure 7: Rltpr is essential for CD28-induced translocation of PKC-θ and CARMA1 to the cSMAC.
Figure 8: RltprBas enhances endocytosis of CD28.

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Acknowledgements

We thank B. Beutler, D. Ordonez-Rueda, H. Holota, J. Ewbank and R. Lasserre for advice; and G. Kollias for leading the MUGEN European network. Supported by Centre National de la Recherche Scientifique, Institut National de la Santé et de la Recherche Médicale, Agence Nationale de Recherche (ADAPT project to M.M.), European Communities (MASTERSWITCH, SYBILLA and MUGEN projects to B.M.), Precursory Research for Embryonic Science and Technology Program of the Japan Science and Technology Agency (T.Y.), Grant-in-Aid for Scientific Research on Innovative Areas (T.Y.), National Science Foundation (NSF 1121793 to C.W.), Groupe d'Intérêt Scientifique–Infrastructures en Biologie Santé et Agronomie, Centre d'Immunophénomique, the AXA Research Fund (M.C.) and the China Scholarship Council (Y.L.).

Author information

Author notes

  1. Yinming Liang, Margot Cucchetti, Romain Roncagalli and Tadashi Yokosuka: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Marseille, France

    Yinming Liang, Margot Cucchetti, Romain Roncagalli, Aurélie Malzac, Elodie Bertosio, Bernard Malissen & Marie Malissen

  2. Institut National de la Santé et de la Recherche Médicale U1104, Marseille, France

    Yinming Liang, Margot Cucchetti, Romain Roncagalli, Aurélie Malzac, Elodie Bertosio, Bernard Malissen & Marie Malissen

  3. Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7280, Marseille, France.,

    Yinming Liang, Margot Cucchetti, Romain Roncagalli, Aurélie Malzac, Elodie Bertosio, Bernard Malissen & Marie Malissen

  4. Laboratory for Cell Signaling, RIKEN Research Center for Allergy and Immunology, Yokohama, Japan

    Tadashi Yokosuka & Takashi Saito

  5. Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, Saitama, Japan

    Tadashi Yokosuka

  6. Aix-Marseille Université, Technologie Avancée pour le Génome et la Clinique, Unité Mixte de Recherche S1090, Marseille, France.,

    Jean Imbert

  7. Institut National de la Santé et de la Recherche Médicale U1090, Marseille, France

    Jean Imbert

  8. Genome Biology and Bioinformatics Group, Hubrecht Institute, Utrecht, The Netherlands

    Isaac J Nijman

  9. Exbio Praha, Vestec, Czech Republic

    Miloslav Suchanek

  10. World Premier International Research Center, Immunology Frontier Center, Osaka University, Osaka, Japan

    Takashi Saito

  11. School of Cell and Molecular Medicine, University of Bristol, Bristol, UK

    Christoph Wülfing

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  1. Yinming Liang
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Contributions

M.M. and B.M. conceived of the project; Y.L. and M.M. designed and did the experiments for Figures 1,2,3,4,5 and 8 and Supplementary Figures 1, 3 and 5; M.C. and R.R. designed and did the experiments for Supplementary Figures 2 and 4; C.W. designed and did the experiments for Supplementary Figure 7 and Supplementary Movies 1 and 2; T.Y., M.C. and T.S. designed and did the experiments for Figures 6 and 7 and Supplementary Figure 6; Y.L., J.I. and I.J.N. contributed to the next-generation sequencing; J.I. did the bioinformatic analysis; A.M., E.B. and M.S. contributed reagents and technical support; and M.M. and B.M. wrote the manuscript.

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Correspondence to Bernard Malissen.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 (PDF 3060 kb)

Supplementary Movie 1

Representative interaction of 5C.C7 CD4+ T cells retrovirally transduced to express the Rltpr-YPET sensor with MCC-pulsed mouse CH27 B lymphoma cells. DIC images are shown on the top, with matching top-down, maximum projections of 3D sensor fluorescence data shown at the bottom. The sensor fluorescence intensity is displayed in a rainbow-like, false-color scale (increasing from blue to red). (MOV 753 kb)

Supplementary Movie 2

Representative interaction of 5C.C7 CD4+ T cells retrovirally transduced to express the RltprBas-YPET sensor with MCC-pulsed mouse CH27 B lymphoma cells. DIC images are shown on the top, with matching top-down, maximum projections of 3D sensor fluorescence data shown at the bottom. The sensor fluorescence intensity is displayed in a rainbow-like, false-color scale (increasing from blue to red). (MOV 824 kb)

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Liang, Y., Cucchetti, M., Roncagalli, R. et al. The lymphoid lineage–specific actin-uncapping protein Rltpr is essential for costimulation via CD28 and the development of regulatory T cells. Nat Immunol 14, 858–866 (2013). https://doi.org/10.1038/ni.2634

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