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Inhibition of type 1 diabetes by upregulation of the circadian rhythm-related aryl hydrocarbon receptor nuclear translocator-like 2

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Abstract

The genetic locus Idd6 is involved in type 1 diabetes development in the non-obese diabetic (NOD) mouse through its effect on the immune system and in particular, on T cell activities. Analysis of congenic strains for Idd6 has established the Aryl hydrocarbon receptor nuclear translocator-like 2 (Arntl2) as a likely candidate gene. In this study we investigate the role of Arntl2 in the autoimmune disease and T cell activation. An Arntl2 expressing plasmid was transfected into CD4+ T cells by nucleofection. Expression levels of cytokines and CD4+ T cell activation markers, cell death, apoptosis, and cell proliferation rates were characterized in ex vivo experiments whilst in vivo the transfected cells were transferred into NOD.SCID mice to monitor diabetes development. The results demonstrate that Arntl2 overexpression leads to inhibition of CD4+ T cell proliferation and decreases in their diabetogenic activity without influence on the expression levels of cytokines, CD4+ T cell activation markers, cell death, and apoptosis. Our findings suggest that Arntl2 at the Idd6 locus may act via the inhibition of CD4+ T cell proliferation and the reduction in the diabetogenic activity of CD4+ T cells to protect against autoimmune type 1 diabetes in the NOD mice.

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Abbreviations

CFSE:

Carboxyfluorescein diacetate succinimidyl ester

Arntl2:

Aryl hydrocarbon receptor nuclear translocator-like 2

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Acknowledgements

We thank Joëlle Morin and Xiaoming Zhang for their excellent technical assistance. This work was supported by grants from the ANR (ANR-06-PHYSIO-016-01), the ARD, and the EFSD/JDRF/Novo Nordisk Programme and by recurrent funding from the CNRS, INSERM, and Institut Pasteur.

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Authors and Affiliations

  1. Institut Pasteur, Unité de Génétique Moléculaire Murine, 25 rue du Docteur Roux, 75724, Paris Cedex 15, France

    Chen-Xia He, Philip Avner & Ute C. Rogner

  2. CNRS URA 2578, 25 rue du Docteur Roux, 75724, Paris Cedex 15, France

    Chen-Xia He, Philip Avner & Ute C. Rogner

  3. INSERM U561, Hôpital Saint Vincent de Paul, 82 avenue Denfert Rochereau, 75014, Paris, France

    Nicolas Prevot & Christian Boitard

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  1. Chen-Xia He
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  2. Nicolas Prevot
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  3. Christian Boitard
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  4. Philip Avner
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  5. Ute C. Rogner
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Correspondence to Ute C. Rogner.

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Fig. S1

Arntl2 is without influence on the mRNA levels of cytokines IL-2, IL-4, IL-10, and IFN-γ in CD4+ T cells. a Q-RT-PCR assays showing that the expression levels of cytokines were comparable in splenic CD4+ T cells from NOD control mice and the congenic strain 6.VIII. Samples in each group were from six to eight females of 6–8 weeks of age. P > 0.05 for IL-2, IL-4, and IL-10; P = 0.03 for IFN-γ. b Arntl2 overexpression (24 h after transfection) in CD4+ T cells ex vivo failed to change the mRNA levels of the cytokines under non-stimulation conditions (n = 3; P > 0.05). (PDF 28 kb)

Fig. S2

Expression levels of cellular markers were similar in CD4+ T cells transfected with control plasmid and Arntl2 expressing plasmid (24 h after transfection). FACS histograms show the counts (Y-axis) and fluorescence intensities (X-axis) for anti-CD25, anti-CD62L, anti-CD44, anti-CD69, anti-GITR, anti-ICOS, and anti-TLR1 labeled cells. Culture control is the cell control that was not treated with the transfection process but only cultured in parallel. Transfection control is the cell control that was treated with the transfection process without adding plasmid. Gated events were between 15,000 and 25,000. (PDF 66 kb)

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He, CX., Prevot, N., Boitard, C. et al. Inhibition of type 1 diabetes by upregulation of the circadian rhythm-related aryl hydrocarbon receptor nuclear translocator-like 2. Immunogenetics 62, 585–592 (2010). https://doi.org/10.1007/s00251-010-0467-7

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  • DOI: https://doi.org/10.1007/s00251-010-0467-7

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