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Stability and function of regulatory T cells is maintained by a neuropilin-1–semaphorin-4a axis

Nature volume 501, pages 252–256 (2013)Cite this article

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Abstract

Regulatory T cells (Treg cells) have a crucial role in the immune system by preventing autoimmunity, limiting immunopathology, and maintaining immune homeostasis1. However, they also represent a major barrier to effective anti-tumour immunity and sterilizing immunity to chronic viral infections1. The transcription factor Foxp3 has a major role in the development and programming of Treg cells2,3. The relative stability of Treg cells at inflammatory disease sites has been a highly contentious subject4,5,6. There is considerable interest in identifying pathways that control the stability of Treg cells as many immune-mediated diseases are characterized by either exacerbated or limited Treg-cell function. Here we show that the immune-cell-expressed ligand semaphorin-4a (Sema4a) and the Treg-cell-expressed receptor neuropilin-1 (Nrp1) interact both in vitro, to potentiate Treg-cell function and survival, and in vivo, at inflammatory sites. Using mice with a Treg-cell-restricted deletion of Nrp1, we show that Nrp1 is dispensable for suppression of autoimmunity and maintenance of immune homeostasis, but is required by Treg cells to limit anti-tumour immune responses and to cure established inflammatory colitis. Sema4a ligation of Nrp1 restrained Akt phosphorylation cellularly and at the immunologic synapse by phosphatase and tensin homologue (PTEN), which increased nuclear localization of the transcription factor Foxo3a. The Nrp1-induced transcriptome promoted Treg-cell stability by enhancing quiescence and survival factors while inhibiting programs that promote differentiation. Importantly, this Nrp1-dependent molecular program is evident in intra-tumoral Treg cells. Our data support a model in which Treg-cell stability can be subverted in certain inflammatory sites, but is maintained by a Sema4a–Nrp1 axis, highlighting this pathway as a potential therapeutic target that could limit Treg-cell-mediated tumour-induced tolerance without inducing autoimmunity.

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Figure 1: Sema4a binds Nrp1 to potentiate Treg-cell function and survival in vitro.
Figure 2: Nrp1-deficient Treg cells fail to suppress anti-tumour immune responses.
Figure 3: Ligation of Nrp1 by Sema4a promotes Treg-cell stability through modulation of Akt–mTOR signalling.
Figure 4: Tumour-infiltrating Treg cells bear a signature similar to Sema4a–Nrp1 ligation.

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Gene Expression Omnibus

Data deposits

The data discussed in this publication have been deposited in the NCBI Gene Expression Omnibus and are accessible through GEO Series accession number GSE41185.

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Acknowledgements

The authors would like to thank E. J. Wherry and H. Chi for advice; A. Rudensky, D. Cheresh, K. Yang, T. Geiger and H. Chi for mice; D. R. Green for plasmids; and B. Triplett, M. Howard and M. McKenna at St Louis Cord Blood Bank for cord blood samples. The authors also thank K. Forbes and A. McKenna for maintenance, breeding and genotyping of mouse colonies; A. Castellaw for preparation of human cord blood samples; K. M. Vignali for assistance with cloning; A. Herrada for generating Nrp1-IgG1; A. L. Szymczak-Workman for assistance with histological analysis; S. Morgan, G. Lennon and R. Cross of the St Jude Immunology Flow Lab for cell sorting; the staff of the Shared Animal Resource Center at St Jude Children's Research Hospital for the animal husbandry; the Hartwell Center for Biotechnology and Bioinformatics at St Jude Children's Research Hospital for Affymetrix microarray analysis; the Veterinary Pathology Core for histological preparation; and the Immunology Department at St Jude Children's Research Hospital for helpful discussions. This work was supported by the National Institutes of Health (R01 AI091977 and AI039480 to D.A.A.V.; F32 AI098383 to G.M.D.), NCI Comprehensive Cancer Center Support CORE grant (CA21765, to D.A.A.V.) and ALSAC (to D.A.A.V.).

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Author notes

  1. Greg M. Delgoffe and Seng-Ryong Woo: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Immunology, St Jude Children's Research Hospital, Memphis, 38105, Tennessee, USA

    Greg M. Delgoffe, Seng-Ryong Woo, Meghan E. Turnis, David M. Gravano, Cliff Guy, Abigail E. Overacre, Matthew L. Bettini, Creg J. Workman & Dario A. A. Vignali

  2. Integrated Biomedical Sciences Program, University of Tennessee Health Science Center, Memphis, 38163, Tennessee, USA

    Abigail E. Overacre

  3. Department of Pathology, St Jude Children's Research Hospital, Memphis, 38105, Tennessee, USA

    Peter Vogel

  4. Computational Biology, St Jude Children's Research Hospital, Memphis, 38105, Tennessee, USA

    David Finkelstein

  5. R&D Systems Inc., Minneapolis, 55413, Minnesota, USA

    Jody Bonnevier

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Contributions

G.M.D. designed and performed most of the experiments and wrote the manuscript. S.-R.W. performed critical initial experiments and identified Sema4a and Nrp1 as the ligand–receptor pair. M.E.T. conducted many of the tumour experiments. D.M.G. performed a substantial portion of the colitis experiments. C.G. performed TIRF microscopy. M.L.B. assisted with the Foxp3-deficiency rescue experiments. A.E.O. assisted with several experiments. P.V. performed histological analysis. D.F. performed computational analysis of the microarray data. J.B. provided the blocking monoclonal antibodies to Sema4a and Nrp1. C.J.W. conducted and curated the initial microarray analysis. D.A.A.V. conceived the project, directed the research and wrote the manuscript. All authors edited and approved the manuscript.

Corresponding author

Correspondence to Dario A. A. Vignali.

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Competing interests

J. Bonnevier is an employee of R&D Systems.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-15. (PDF 1424 kb)

Supplementary Table 1

This file contains geneset enrichment analysis for Nrp1-upregulated genesets. (XLS 44 kb)

Supplementary Table 2

This file contains geneset enrichment analysis for Nrp1-downregulated genesets. (XLS 167 kb)

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Delgoffe, G., Woo, SR., Turnis, M. et al. Stability and function of regulatory T cells is maintained by a neuropilin-1–semaphorin-4a axis. Nature 501, 252–256 (2013). https://doi.org/10.1038/nature12428

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