Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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

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.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

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.

Accession codes

Accessions

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.

References

  1. Vignali, D. A., Collison, L. W. & Workman, C. J. How regulatory T cells work. Nature Rev. Immunol. 8, 523–532 (2008)

    CAS  Article  Google Scholar 

  2. Fontenot, J. D., Gavin, M. A. & Rudensky, A. Y. Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nature Immunol. 4, 330–336 (2003)

    CAS  Article  Google Scholar 

  3. Hori, S., Nomura, T. & Sakaguchi, S. Control of regulatory T cell development by the transcription factor Foxp3. Science 299, 1057–1061 (2003)

    CAS  ADS  Article  Google Scholar 

  4. Miyao, T. et al. Plasticity of Foxp3+ T cells reflects promiscuous Foxp3 expression in conventional T cells but not reprogramming of regulatory T cells. Immunity 36, 262–275 (2012)

    CAS  Article  Google Scholar 

  5. Rubtsov, Y. P. et al. Stability of the regulatory T cell lineage in vivo. Science 329, 1667–1671 (2010)

    CAS  ADS  Article  Google Scholar 

  6. Zhou, X. et al. Instability of the transcription factor Foxp3 leads to the generation of pathogenic memory T cells in vivo. Nature Immunol. 10, 1000–1007 (2009)

    CAS  ADS  Article  Google Scholar 

  7. Qiao, M., Thornton, A. M. & Shevach, E. M. CD4+ CD25+ regulatory T cells render naive CD4+ CD25- T cells anergic and suppressive. Immunology 120, 447–455 (2007); corrigendum. 121, 146 (2007)

    CAS  Article  Google Scholar 

  8. Collison, L. W., Pillai, M. R., Chaturvedi, V. & Vignali, D. A. Regulatory T cell suppression is potentiated by target T cells in a cell contact, IL-35- and IL-10-dependent manner. J. Immunol. 182, 6121–6128 (2009)

    CAS  Article  Google Scholar 

  9. Kumanogoh, A. et al. Class IV semaphorin Sema4A enhances T-cell activation and interacts with Tim-2. Nature 419, 629–633 (2002)

    CAS  ADS  Article  Google Scholar 

  10. Bruder, D. et al. Neuropilin-1: a surface marker of regulatory T cells. Eur. J. Immunol. 34, 623–630 (2004)

    CAS  Article  Google Scholar 

  11. Kolodkin, A. L. et al. Neuropilin is a semaphorin III receptor. Cell 90, 753–762 (1997)

    CAS  Article  Google Scholar 

  12. Weiss, J. M. et al. Neuropilin 1 is expressed on thymus-derived natural regulatory T cells, but not mucosa-generated induced Foxp3+ T reg cells. J. Exp. Med. 209, 1723–1742 (2012)

    CAS  Article  Google Scholar 

  13. Yadav, M. et al. Neuropilin-1 distinguishes natural and inducible regulatory T cells among regulatory T cell subsets in vivo. J. Exp. Med. 209, 1713–1722 (2012)

    CAS  Article  Google Scholar 

  14. Milpied, P. et al. Neuropilin-1 is not a marker of human Foxp3+ Treg. Eur. J. Immunol. 39, 1466–1471 (2009)

    CAS  Article  Google Scholar 

  15. Piechnik, A. et al. The VEGF receptor neuropilin-1 NRP1) represents a promising novel target for chronic lymphocytic leukemia patients. Int. J. Cancer http://dx.doi.org/10.1002/ijc.28135 (28 February 2013)

  16. Nishikawa, H. & Sakaguchi, S. Regulatory T cells in tumor immunity. Int. J. Cancer 127, 759–767 (2010)

    CAS  Google Scholar 

  17. Kim, J. M., Rasmussen, J. P. & Rudensky, A. Y. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice. Nature Immunol. 8, 191–197 (2007)

    CAS  Article  Google Scholar 

  18. Wilde, S. et al. Human antitumor CD8+ T cells producing Th1 polycytokines show superior antigen sensitivity and tumor recognition. J. Immunol. 189, 598–605 (2012)

    CAS  Article  Google Scholar 

  19. Demoulin, S., Herfs, M., Delvenne, P. & Hubert, P. Tumor microenvironment converts plasmacytoid dendritic cells into immunosuppressive/tolerogenic cells: insight into the molecular mechanisms. J. Leukoc. Biol. 93, 343–352 (2013)

    CAS  Article  Google Scholar 

  20. Castro-Rivera, E., Ran, S., Brekken, R. A. & Minna, J. D. Semaphorin 3B inhibits the phosphatidylinositol 3-kinase/Akt pathway through neuropilin-1 in lung and breast cancer cells. Cancer Res. 68, 8295–8303 (2008)

    CAS  Article  Google Scholar 

  21. Crellin, N. K., Garcia, R. V. & Levings, M. K. Altered activation of AKT is required for the suppressive function of human CD4+CD25+ T regulatory cells. Blood 109, 2014–2022 (2007)

    CAS  Article  Google Scholar 

  22. Haxhinasto, S., Mathis, D. & Benoist, C. The AKT-mTOR axis regulates de novo differentiation of CD4+Foxp3+ cells. J. Exp. Med. 205, 565–574 (2008)

    CAS  Article  Google Scholar 

  23. Franke, T. F. et al. The protein kinase encoded by the Akt proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase. Cell 81, 727–736 (1995)

    CAS  Article  Google Scholar 

  24. Stambolic, V. et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell 95, 29–39 (1998)

    CAS  Article  Google Scholar 

  25. Pellet-Many, C., Frankel, P., Jia, H. & Zachary, I. Neuropilins: structure, function and role in disease. Biochem. J. 411, 211–226 (2008)

    CAS  Article  Google Scholar 

  26. Merkenschlager, M. & von Boehmer, H. PI3 kinase signalling blocks Foxp3 expression by sequestering Foxo factors. J. Exp. Med. 207, 1347–1350 (2010)

    CAS  Article  Google Scholar 

  27. Kerdiles, Y. M. et al. Foxo transcription factors control regulatory T cell development and function. Immunity 33, 890–904 (2010); erratum. 34, 135 (2011)

    CAS  Article  Google Scholar 

  28. Ouyang, W. et al. Foxo proteins cooperatively control the differentiation of Foxp3+ regulatory T cells. Nature Immunol. 11, 618–627 (2010)

    CAS  Article  Google Scholar 

  29. Heng, T. S. & Painter, M. W. The Immunological Genome Project: networks of gene expression in immune cells. Nature Immunol. 9, 1091–1094 (2008)

    CAS  Article  Google Scholar 

  30. Sawant, A. et al. Depletion of plasmacytoid dendritic cells inhibits tumor growth and prevents bone metastasis of breast cancer cells. J. Immunol. 189, 4258–4265 (2012)

    CAS  Article  Google Scholar 

  31. Guy, C. S. et al. Distinct TCR signaling pathways drive proliferation and cytokine production in T cells. Nature Immunol. 14, 262–270 (2013)

    CAS  Article  Google Scholar 

Download references

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.).

Author information

Authors and Affiliations

Authors

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.

Ethics declarations

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)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature12428

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing