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USP21 prevents the generation of T-helper-1-like Treg cells

View Article: PubMed Central - PubMed

ABSTRACT

FOXP3+ Regulatory T (Treg) cells play a key role in the maintenance of immune homeostasis and tolerance. Disruption of Foxp3 expression results in the generation of instable Treg cells and acquisition of effector T-cell-like function. Here we report that the E3 deubiquitinase USP21 prevents the depletion of FOXP3 at the protein level and restricts the generation of T-helper-1-like Treg cells. Mice depleted of Usp21 specifically in Treg cells display immune disorders characterized by spontaneous T-cell activation and excessive T-helper type 1 (Th1) skewing of Treg cells into Th1-like Treg cells. USP21 stabilizes FOXP3 protein by mediating its deubiquitination and maintains the expression of Treg signature genes. Our results demonstrate how USP21 prevents FOXP3 protein depletion and controls Treg lineage stability in vivo.

No MeSH data available.


Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms.(a) Clinical severity of EAE in WT (n=8) and KO (n=7) mice was monitored for 27 days after immunization with MOG peptide. Statistical analysis of the clinical signs was further described in Table 1. (b) Representative figure shown the expression of IFN-γ and IL-17A by CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice on day 27 post EAE induction. (c) Percentage of IFN-γ+ and IL-17A+ cells among CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice as indicated in b. (d) CD45.2+ WT Treg or USP21-ΔTreg cells were transferred intravenously into CD45.1 mice at the onset of EAE (day 12). At the peak of EAE (day 17), the transferred CD45.2+ Treg cells were analysed for FOXP3 expression in the CNS from the CD45.1 mice. (e) Mean fluorescent intensity (MFI) of FOXP3 among transferred CD45.2+ Treg cells as indicated in e. (n=5 for each group). All data represent means±s.d. *P≤0.05, **P≤0.01, ***P≤0.001, as determined by Student's t-test. NS, not significant.
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f6: Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms.(a) Clinical severity of EAE in WT (n=8) and KO (n=7) mice was monitored for 27 days after immunization with MOG peptide. Statistical analysis of the clinical signs was further described in Table 1. (b) Representative figure shown the expression of IFN-γ and IL-17A by CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice on day 27 post EAE induction. (c) Percentage of IFN-γ+ and IL-17A+ cells among CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice as indicated in b. (d) CD45.2+ WT Treg or USP21-ΔTreg cells were transferred intravenously into CD45.1 mice at the onset of EAE (day 12). At the peak of EAE (day 17), the transferred CD45.2+ Treg cells were analysed for FOXP3 expression in the CNS from the CD45.1 mice. (e) Mean fluorescent intensity (MFI) of FOXP3 among transferred CD45.2+ Treg cells as indicated in e. (n=5 for each group). All data represent means±s.d. *P≤0.05, **P≤0.01, ***P≤0.001, as determined by Student's t-test. NS, not significant.

Mentions: We further challenged Usp21fl/flFoxp3Cre mice with MOG peptide in an experimental allergic encephalomyelitis (EAE) disease model. We found that Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms and failed to restrict late stages of disease development (Fig. 6a; Table 1). Disease severity was accompanied by increased production of IFN-γ and IL-17A in CD4+YFP− effector T cells from Usp21fl/flFoxp3Cre mice (Fig. 6b,c). We then transferred CD45.2+ WT Treg or USP21-ΔTreg cells into EAE-bearing CD45.1 mice, and CD45.2+ USP21-ΔTreg cells became instable through FOXP3 loss (Fig. 6d,e). Therefore, USP21-ΔTreg cells fail to limit EAE disease development at later phases of disease progression, possibly due to the loss of FOXP3 expression and suppressive function.


USP21 prevents the generation of T-helper-1-like Treg cells
Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms.(a) Clinical severity of EAE in WT (n=8) and KO (n=7) mice was monitored for 27 days after immunization with MOG peptide. Statistical analysis of the clinical signs was further described in Table 1. (b) Representative figure shown the expression of IFN-γ and IL-17A by CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice on day 27 post EAE induction. (c) Percentage of IFN-γ+ and IL-17A+ cells among CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice as indicated in b. (d) CD45.2+ WT Treg or USP21-ΔTreg cells were transferred intravenously into CD45.1 mice at the onset of EAE (day 12). At the peak of EAE (day 17), the transferred CD45.2+ Treg cells were analysed for FOXP3 expression in the CNS from the CD45.1 mice. (e) Mean fluorescent intensity (MFI) of FOXP3 among transferred CD45.2+ Treg cells as indicated in e. (n=5 for each group). All data represent means±s.d. *P≤0.05, **P≤0.01, ***P≤0.001, as determined by Student's t-test. NS, not significant.
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f6: Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms.(a) Clinical severity of EAE in WT (n=8) and KO (n=7) mice was monitored for 27 days after immunization with MOG peptide. Statistical analysis of the clinical signs was further described in Table 1. (b) Representative figure shown the expression of IFN-γ and IL-17A by CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice on day 27 post EAE induction. (c) Percentage of IFN-γ+ and IL-17A+ cells among CD4+YFP− effector T cells in the spleen, peripheral lymph nodes and CNS from WT (n=5) and KO (n=5) mice as indicated in b. (d) CD45.2+ WT Treg or USP21-ΔTreg cells were transferred intravenously into CD45.1 mice at the onset of EAE (day 12). At the peak of EAE (day 17), the transferred CD45.2+ Treg cells were analysed for FOXP3 expression in the CNS from the CD45.1 mice. (e) Mean fluorescent intensity (MFI) of FOXP3 among transferred CD45.2+ Treg cells as indicated in e. (n=5 for each group). All data represent means±s.d. *P≤0.05, **P≤0.01, ***P≤0.001, as determined by Student's t-test. NS, not significant.
Mentions: We further challenged Usp21fl/flFoxp3Cre mice with MOG peptide in an experimental allergic encephalomyelitis (EAE) disease model. We found that Usp21fl/flFoxp3Cre mice developed more severe EAE symptoms and failed to restrict late stages of disease development (Fig. 6a; Table 1). Disease severity was accompanied by increased production of IFN-γ and IL-17A in CD4+YFP− effector T cells from Usp21fl/flFoxp3Cre mice (Fig. 6b,c). We then transferred CD45.2+ WT Treg or USP21-ΔTreg cells into EAE-bearing CD45.1 mice, and CD45.2+ USP21-ΔTreg cells became instable through FOXP3 loss (Fig. 6d,e). Therefore, USP21-ΔTreg cells fail to limit EAE disease development at later phases of disease progression, possibly due to the loss of FOXP3 expression and suppressive function.

View Article: PubMed Central - PubMed

ABSTRACT

FOXP3+ Regulatory T (Treg) cells play a key role in the maintenance of immune homeostasis and tolerance. Disruption of Foxp3 expression results in the generation of instable Treg cells and acquisition of effector T-cell-like function. Here we report that the E3 deubiquitinase USP21 prevents the depletion of FOXP3 at the protein level and restricts the generation of T-helper-1-like Treg cells. Mice depleted of Usp21 specifically in Treg cells display immune disorders characterized by spontaneous T-cell activation and excessive T-helper type 1 (Th1) skewing of Treg cells into Th1-like Treg cells. USP21 stabilizes FOXP3 protein by mediating its deubiquitination and maintains the expression of Treg signature genes. Our results demonstrate how USP21 prevents FOXP3 protein depletion and controls Treg lineage stability in vivo.

No MeSH data available.