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The nuclear orphan receptor Nr4a2 induces Foxp3 and regulates differentiation of CD4+ T cells.

Sekiya T, Kashiwagi I, Inoue N, Morita R, Hori S, Waldmann H, Rudensky AY, Ichinose H, Metzger D, Chambon P, Yoshimura A - Nat Commun (2011)

Bottom Line: Regulatory T cells (Tregs) have a central role in maintaining immune homoeostasis through various mechanisms.Nr4a2 binds to regulatory regions of Foxp3, where it mediates permissive histone modifications.Nr4a2-deficeint Tregs are prone to lose Foxp3 expression and have attenuated suppressive ability both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Keio University School of Medicine, Tokyo 160-8582, Japan.

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Nr4a families vary in their ability to induce Foxp3 and to repress IFN-γ.(a) Top row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on Foxp3 expression. Bottom row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on IFN-γ expression. eMIGR or eMIGR-Nr4a retrovirus-transduced cells were restimulated with phorbol myristate acetate (PMA) and ionomycine at 60 h after infection for 8 h. (b, c) Quantification of the results of a. (d) Effects of Nr4a2 transductions on Foxp3 expression (top row) and IFN-γ repression (bottom row) under Th1 and Th17 conditions. (e) Quantification of the results of the eMIGR-Nr4a2 retrovirus-transduced cells in d. (f) Schematic representation of Nr4a2, Nr4a3 and the chimeric protein of Nr4a3 in which the N-terminal of Nr4a3 is replaced with that of Nr4a2 (Nr4a2-a3 chimera). DBD, DNA-binding domain; LBD, ligand-binding domain. (g) Foxp3 induction (top row) and IFN-γ repression (bottom row) by Nr4a2, Nr4a3 and Nr4a2-a3 chimera. Cells were transduced with eMIGR1- or eMIGR1-Nr4a-retorviruses, and Foxp3 induction and IFN-γ repression were analysed. (h) Transcriptional activities as monomers of Nr4a2, Nr4a3 and Nr4a2-a3 chimera were demonstrated using NBRE-luc in 293T cells. (i) Quantification of the result in g. Numbers in FACS plots represent percentages of CD4+ T cells in the gated area. Data in (b, c, e, h, i) are representative of three independent experiments (mean and s.d. of triplicate). *P< 0.05 and **P<0.01 (two-tailed Student's t-test).
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f3: Nr4a families vary in their ability to induce Foxp3 and to repress IFN-γ.(a) Top row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on Foxp3 expression. Bottom row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on IFN-γ expression. eMIGR or eMIGR-Nr4a retrovirus-transduced cells were restimulated with phorbol myristate acetate (PMA) and ionomycine at 60 h after infection for 8 h. (b, c) Quantification of the results of a. (d) Effects of Nr4a2 transductions on Foxp3 expression (top row) and IFN-γ repression (bottom row) under Th1 and Th17 conditions. (e) Quantification of the results of the eMIGR-Nr4a2 retrovirus-transduced cells in d. (f) Schematic representation of Nr4a2, Nr4a3 and the chimeric protein of Nr4a3 in which the N-terminal of Nr4a3 is replaced with that of Nr4a2 (Nr4a2-a3 chimera). DBD, DNA-binding domain; LBD, ligand-binding domain. (g) Foxp3 induction (top row) and IFN-γ repression (bottom row) by Nr4a2, Nr4a3 and Nr4a2-a3 chimera. Cells were transduced with eMIGR1- or eMIGR1-Nr4a-retorviruses, and Foxp3 induction and IFN-γ repression were analysed. (h) Transcriptional activities as monomers of Nr4a2, Nr4a3 and Nr4a2-a3 chimera were demonstrated using NBRE-luc in 293T cells. (i) Quantification of the result in g. Numbers in FACS plots represent percentages of CD4+ T cells in the gated area. Data in (b, c, e, h, i) are representative of three independent experiments (mean and s.d. of triplicate). *P< 0.05 and **P<0.01 (two-tailed Student's t-test).

Mentions: Because Nr4a mediates various physiological events, some in a redundant manner and others in a non-redundant manner, we compared the ability of the three family members to induce Foxp3. Importantly, both Nr4a1 and Nr4a3 have also been reported to be abundant in Tregs31 and inducible by TCR stimulation32. We found that Nr4a1 rarely induced Foxp3, whereas Nr4a3 did so to an intermediate degree (Fig. 3a,b), despite the expression levels of these proteins were similar (Supplementary Fig. S3a).


The nuclear orphan receptor Nr4a2 induces Foxp3 and regulates differentiation of CD4+ T cells.

Sekiya T, Kashiwagi I, Inoue N, Morita R, Hori S, Waldmann H, Rudensky AY, Ichinose H, Metzger D, Chambon P, Yoshimura A - Nat Commun (2011)

Nr4a families vary in their ability to induce Foxp3 and to repress IFN-γ.(a) Top row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on Foxp3 expression. Bottom row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on IFN-γ expression. eMIGR or eMIGR-Nr4a retrovirus-transduced cells were restimulated with phorbol myristate acetate (PMA) and ionomycine at 60 h after infection for 8 h. (b, c) Quantification of the results of a. (d) Effects of Nr4a2 transductions on Foxp3 expression (top row) and IFN-γ repression (bottom row) under Th1 and Th17 conditions. (e) Quantification of the results of the eMIGR-Nr4a2 retrovirus-transduced cells in d. (f) Schematic representation of Nr4a2, Nr4a3 and the chimeric protein of Nr4a3 in which the N-terminal of Nr4a3 is replaced with that of Nr4a2 (Nr4a2-a3 chimera). DBD, DNA-binding domain; LBD, ligand-binding domain. (g) Foxp3 induction (top row) and IFN-γ repression (bottom row) by Nr4a2, Nr4a3 and Nr4a2-a3 chimera. Cells were transduced with eMIGR1- or eMIGR1-Nr4a-retorviruses, and Foxp3 induction and IFN-γ repression were analysed. (h) Transcriptional activities as monomers of Nr4a2, Nr4a3 and Nr4a2-a3 chimera were demonstrated using NBRE-luc in 293T cells. (i) Quantification of the result in g. Numbers in FACS plots represent percentages of CD4+ T cells in the gated area. Data in (b, c, e, h, i) are representative of three independent experiments (mean and s.d. of triplicate). *P< 0.05 and **P<0.01 (two-tailed Student's t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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f3: Nr4a families vary in their ability to induce Foxp3 and to repress IFN-γ.(a) Top row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on Foxp3 expression. Bottom row: effects of Nr4a1, Nr4a2 and Nr4a3 expression on IFN-γ expression. eMIGR or eMIGR-Nr4a retrovirus-transduced cells were restimulated with phorbol myristate acetate (PMA) and ionomycine at 60 h after infection for 8 h. (b, c) Quantification of the results of a. (d) Effects of Nr4a2 transductions on Foxp3 expression (top row) and IFN-γ repression (bottom row) under Th1 and Th17 conditions. (e) Quantification of the results of the eMIGR-Nr4a2 retrovirus-transduced cells in d. (f) Schematic representation of Nr4a2, Nr4a3 and the chimeric protein of Nr4a3 in which the N-terminal of Nr4a3 is replaced with that of Nr4a2 (Nr4a2-a3 chimera). DBD, DNA-binding domain; LBD, ligand-binding domain. (g) Foxp3 induction (top row) and IFN-γ repression (bottom row) by Nr4a2, Nr4a3 and Nr4a2-a3 chimera. Cells were transduced with eMIGR1- or eMIGR1-Nr4a-retorviruses, and Foxp3 induction and IFN-γ repression were analysed. (h) Transcriptional activities as monomers of Nr4a2, Nr4a3 and Nr4a2-a3 chimera were demonstrated using NBRE-luc in 293T cells. (i) Quantification of the result in g. Numbers in FACS plots represent percentages of CD4+ T cells in the gated area. Data in (b, c, e, h, i) are representative of three independent experiments (mean and s.d. of triplicate). *P< 0.05 and **P<0.01 (two-tailed Student's t-test).
Mentions: Because Nr4a mediates various physiological events, some in a redundant manner and others in a non-redundant manner, we compared the ability of the three family members to induce Foxp3. Importantly, both Nr4a1 and Nr4a3 have also been reported to be abundant in Tregs31 and inducible by TCR stimulation32. We found that Nr4a1 rarely induced Foxp3, whereas Nr4a3 did so to an intermediate degree (Fig. 3a,b), despite the expression levels of these proteins were similar (Supplementary Fig. S3a).

Bottom Line: Regulatory T cells (Tregs) have a central role in maintaining immune homoeostasis through various mechanisms.Nr4a2 binds to regulatory regions of Foxp3, where it mediates permissive histone modifications.Nr4a2-deficeint Tregs are prone to lose Foxp3 expression and have attenuated suppressive ability both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Keio University School of Medicine, Tokyo 160-8582, Japan.

Show MeSH
Related in: MedlinePlus