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Interleukin-2 gene variation impairs regulatory T cell function and causes autoimmunity

Nature Genetics volume 39, pages 329–337 (2007)Cite this article

Abstract

Autoimmune diseases are thought to result from imbalances in normal immune physiology and regulation. Here, we show that autoimmune disease susceptibility and resistance alleles on mouse chromosome 3 (Idd3) correlate with differential expression of the key immunoregulatory cytokine interleukin-2 (IL-2). In order to test directly that an approximately twofold reduction in IL-2 underpins the Idd3-linked destabilization of immune homeostasis, we show that engineered haplodeficiency of Il2 gene expression not only reduces T cell IL-2 production by twofold but also mimics the autoimmune dysregulatory effects of the naturally occurring susceptibility alleles of Il2. Reduced IL-2 production achieved by either genetic mechanism correlates with reduced function of CD4+ CD25+ regulatory T cells, which are critical for maintaining immune homeostasis.

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Figure 1: Frequency of diabetes in Idd3-congenic NOD strains.
Figure 2: Correlation of Idd3-linked disease susceptibility and resistance with Il2 expression.
Figure 3: Development and function of IGRP206–214-reactive CD8+ T cells in the presence or absence of the B6 Idd3 region.
Figure 4: IL-2 production competency and diabetes susceptibility of Il2-hemizygous mice.
Figure 5: Enhanced effector function of 8.3-CD8+ CTL in 8.3-NOD.Idd3B6/B6 mice.
Figure 6: Enhanced function of CD4+ CD25+ Treg cells in 8.3-NOD.Idd3B6/B6 mice.
Figure 7: Enhanced function of CD4+ CD25+ Treg cells in 8.3-NOD.Idd3B6/NOD mice is dependent on IL-2 production.

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Acknowledgements

We thank S. Bou, M. Deuma, S. Thiessen and H. Vandermeer for technical assistance with mouse breeding, genotyping and animal care; L. Kennedy and L. Robertson for flow cytometry and A. Shameli, P. Taylor, S. Tsai, U. Walter and Y. Yang for reading the manuscript. NOD.B6 Idd3 and NOD.CZECH Idd3 are available from Taconic through the Emerging Models Program (Lines 1098 and 1590, respectively). This work was supported by grants from the Canadian Institutes of Health Research, the Juvenile Diabetes Research Foundation (JDRF) and the Wellcome Trust. J.Y. was supported by fellowships from the Canadian Diabetes Association and JDRF. L.S.W. and J.A.T. are supported by grants from the JDRF and the Wellcome Trust, and L.S.W. is a JDRF/Wellcome Trust Principal Research Fellow. P. Serra was supported by a studentship from the Alberta Heritage Foundation for Medical Research (AHFMR). P. Santamaria is a Scientist of AHFMR. The JMDRC is supported by the Diabetes Association (Foothills). The availability of NOD congenic mice through the Taconic Farms Emerging Models Program has been supported by grants from the Merck Genome Research Institute, National Institute of Allergy and Infectious Diseases and the JDRF. Requests for materials should be addressed to P. Santamaria (psantama@ucalgary.ca) or L.S.W. (linda.wicker@cimr.cam.ac.uk).

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Author notes

  1. Linda S Wicker and Pere Santamaria: These authors contributed equally to this work.

Authors and Affiliations

  1. Julia McFarlane Diabetes Research Centre (JMDRC) and Department of Microbiology and Infectious Diseases, Institute of Inflammation, Infection and Immunity, Faculty of Medicine, The University of Calgary, Calgary, T2N 4N1, Alberta, Canada

    Jun Yamanouchi, Pau Serra & Pere Santamaria

  2. Department of Medical Genetics, Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 2XY, UK

    Dan Rainbow, Sarah Howlett, Kara Hunter, Valerie E S Garner, Andrea Gonzalez-Munoz, Jan Clark, Riitta Veijola, Paul A Lyons, Barry Healy, Luc J Smink, John A Todd & Linda S Wicker

  3. Merck Research Laboratories, Rahway, 07065, New Jersey, USA

    Rose Cubbon, Show-Ling Chen, Raymond Rosa, Anne Marie Cumiskey & Laurence B Peterson

  4. The Jackson Laboratory, Bar Harbor, 04609, Maine, USA

    David V Serreze

  5. Wellcome Trust Sanger Institute, Hinxton, Cambridge, CB10 1HH, UK

    Simon Gregory & Jane Rogers

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Contributions

J.Y. and P. Santamaria conceived and designed the immunological experiments with transgenic and nontransgenic mice. L.W., J.T. and L.P. conceived and designed the genetic experiments. J.Y., P. Santamaria, L.W. and J.T. wrote the paper. J.Y. and P. Serra carried out the experiments shown in Figures 3a,b, 4,5,6,7 and Supplementary Figures 4,5,6,7. D.R., V.G., S.G., J.R., B.H., L.S. and L.W. did the sequencing and bioinformatic analyses of Idd3 shown in Supplementary Figure 1. S.H., D.R., K.H., V.G., R.R., A.M.C., J.C. and P.L. genotyped the congenic strains and/or generated the RT qPCR data shown in Fig. 2 and 3 and Supplementary Figures 2 and 3. R.C., S.-L.C., P.L., A.G.-M., R.V., L.P. and L.W. produced and monitored the nontransgenic congenic strains described in the paper to generate the survival curves shown in Fig. 1 and the T1D-gene/SNP associations shown in Supplementary Figure 1. D.S. produced the NOD.Il2+/− strain.

Corresponding authors

Correspondence to Linda S Wicker or Pere Santamaria.

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Supplementary information

Supplementary Fig. 1

The Idd3 region of mouse chromosome 3 and sequence coverage track and location of disease-associated SNPs in Il2 and Il21. (PDF 1631 kb)

Supplementary Fig. 2

Correlation of disease susceptibility and resistance with Il2 expression in thymocytes. (PDF 116 kb)

Supplementary Fig. 3

Preferential production of IL-2 pre-mRNA from the resistant Il2 allele occurs in both CD4+ and CD8+ T cells and is a cis-regulated genetic event. (PDF 231 kb)

Supplementary Fig. 4

Phenotype and function of 8.3-CD8+ T cells in the presence or absence of the B6 Idd3 region. (PDF 419 kb)

Supplementary Fig. 5

5 IL-2 production competency as determined by intracellular IL-2 staining. (PDF 531 kb)

Supplementary Fig. 6

Enhanced development and function of CD4+ CD25+ Tregs in nontransgenic NOD.Idd3B6/B6 and B10.BR mice. (PDF 285 kb)

Supplementary Fig. 7

Phenotype and function of CD4+ CD25+ T cells from NOD and NOD.Idd3B6/B6 mice. (PDF 418 kb)

Supplementary Table 1

Primer sequences. (PDF 41 kb)

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Yamanouchi, J., Rainbow, D., Serra, P. et al. Interleukin-2 gene variation impairs regulatory T cell function and causes autoimmunity. Nat Genet 39, 329–337 (2007). https://doi.org/10.1038/ng1958

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