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

A Corrigendum to this article was published on 08 October 2014

This article has been updated

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.

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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.

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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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