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Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells

Nature Medicine volume 19, pages 739–746 (2013)Cite this article

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A Corrigendum to this article was published on 08 October 2014

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Abstract

CD4+ type 1 T regulatory (Tr1) cells are induced in the periphery and have a pivotal role in promoting and maintaining tolerance. The absence of surface markers that uniquely identify Tr1 cells has limited their study and clinical applications. By gene expression profiling of human Tr1 cell clones, we identified the surface markers CD49b and lymphocyte activation gene 3 (LAG-3) as being stably and selectively coexpressed on mouse and human Tr1 cells. We showed the specificity of these markers in mouse models of intestinal inflammation and helminth infection and in the peripheral blood of healthy volunteers. The coexpression of CD49b and LAG-3 enables the isolation of highly suppressive human Tr1 cells from in vitro anergized cultures and allows the tracking of Tr1 cells in the peripheral blood of subjects who developed tolerance after allogeneic hematopoietic stem cell transplantation. The use of these markers makes it feasible to track Tr1 cells in vivo and purify Tr1 cells for cell therapy to induce or restore tolerance in subjects with immune-mediated diseases.

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Figure 1: Identification of CD49b, LAG-3 and CD226 by gene expression profiling of human Tr1 cell clones.
Figure 2: Coexpression of CD49b and LAG-3 identifies human Tr1 cells in vivo in healthy donors.
Figure 3: Coexpression of CD49b and LAG-3 identifies mouse Tr1 cells in mice treated with antibodies to CD3.
Figure 4: In vitro and in vivo regulatory activity of mouse CD4+CD49b+LAG-3+ T cells.
Figure 5: Coexpression of CD49b and LAG-3 is specific for mouse Tr1 cells.
Figure 6: Coexpression of CD49b and LAG-3 allows for the selection of human Tr1 cells in vitro and the enumeration of Tr1 cells in vivo in tolerant subjects.

Change history

  • 14 May 2013

     In the version of this article initially published online, Silvia Gregori’s contribution was incorrectly labeled. She jointly directed the work along with Richard A. Flavell and Maria-Grazia Roncarolo. Additionally, an asterisk in Figure 4c, bottom row, second from right, was incorrectly placed. The errors have been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We wish to thank M. Battaglia (DRI, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute) for helpful scientific discussions and critical revision of the manuscript and we thank L. Passerini (HSR-TIGET, Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute) for providing human CD25bright T cell samples. This work was supported by Telethon Foundation 'Comitato Telethon Fondazione Onlus' Core Grant OSR-TIGET project E2 (Rome) to S.G. and M.-G.R. N.G. was supported by an EMBO long-term postdoctoral fellowship. S.H. was supported by the 'Jung-Stiftung', 'Stiftung Experimentelle Biomedizin' and the Deutsche Forschungsgemeinschaft (DFG) (HU 1714/3-1). R.A.F. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

  1. Nicola Gagliani, Chiara F Magnani, Samuel Huber and Silvia Gregori: These authors contributed equally to this work.

  2. Silvia Gregori, Richard A Flavell and Maria-Grazia Roncarolo: These authors jointly directed this work.

Authors and Affiliations

  1. Division of Regenerative Medicine, San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan, Italy

    Nicola Gagliani, Chiara F Magnani, Monica E Gianolini, Rosa Bacchetta & Maria-Grazia Roncarolo

  2. Department of Immunobiology, School of Medicine, Yale University, New Haven, Connecticut, USA

    Nicola Gagliani, Paula Licona-Limon & Richard A Flavell

  3. I. Medizinische Klinik und Poliklinik, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany

    Samuel Huber & Leonie Brockmann

  4. Department of Public Health and Cell Biology, Tor Vergata University, Rome, Italy

    Monica E Gianolini

  5. Bioflag, Technology Park Polaris, Pula, Italy

    Mauro Pala & Alessandro Bulfone

  6. School of Life Sciences, Peking University, Beijing, China

    Binggege Guo

  7. Division of Experimental Medicine, University of California San Francisco, San Francisco, California, USA

    De'Broski R Herbert

  8. University Centre for Statistics in the Biomedical Sciences (CUSSB), Vita-Salute San Raffaele University, Milan, Italy

    Filippo Trentini & Clelia Di Serio

  9. Laboratorio di Immunogenetica e Biologia dei Trapianti, Fondazione Istituto Mediterraneo di Ematologia (IME), Policlinico Tor Vergata, Rome, Italy

    Marco Andreani

  10. Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut, USA

    Richard A Flavell

  11. Vita-Salute San Raffaele University, Milan, Italy

    Maria-Grazia Roncarolo

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  1. Nicola Gagliani
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Contributions

N.G., S.H. and S.G. designed the experiments, analyzed the data and wrote the manuscript. C.F.M. prepared T cell clones for gene expression profile analysis and performed RT-PCR and FACS analyses on human T cell clones. M.E.G. performed the experiments with human samples. M.P. performed bioinformatics analysis. A.B. supervised microarrays and bioinformatics analysis. H.R.D. and P.L.-L. provided the expertise and supervised the experiments with N. brasiliensis. B.G. performed the mouse RT-PCR. L.B. performed the mouse in vitro suppression assay. F.T. performed statistical analysis on patient samples. C.D.S. supervised statistical analysis on patient samples. R.B. contributed to the selection of candidate markers. M.A. provided the clinical samples. S.G., R.A.F. and M.-G.R. coordinated and supervised the project and wrote the manuscript.

Corresponding authors

Correspondence to Richard A Flavell or Maria-Grazia Roncarolo.

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Gagliani, N., Magnani, C., Huber, S. et al. Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells. Nat Med 19, 739–746 (2013). https://doi.org/10.1038/nm.3179

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