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The Nuclear Orphan Receptor NR2F6 Is a Central Checkpoint for Cancer Immune Surveillance.

Hermann-Kleiter N, Klepsch V, Wallner S, Siegmund K, Klepsch S, Tuzlak S, Villunger A, Kaminski S, Pfeifhofer-Obermair C, Gruber T, Wolf D, Baier G - Cell Rep (2015)

Bottom Line: Nuclear receptor subfamily 2, group F, member 6 (NR2F6) is an orphan member of the nuclear receptor superfamily.Mechanistically, CD4(+) and CD8(+) T cell-intrinsic NR2F6 acts as a direct repressor of the NFAT/AP-1 complex on both the interleukin 2 and the interferon γ cytokine promoters, attenuating their transcriptional thresholds.Altogether, this defines NR2F6 as an intracellular immune checkpoint in effector T cells, governing the amplitude of anti-cancer immunity.

View Article: PubMed Central - PubMed

Affiliation: Translational Cell Genetics, Department for Pharmacology and Genetics, Medical University of Innsbruck, 6020 Innsbruck, Austria. Electronic address: natascha.kleiter@i-med.ac.at.

No MeSH data available.


Related in: MedlinePlus

Nr2f6 Expression Limits Cytokine Secretion of Tumor-Reactive T Cells(A) Cytokine secretion of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) TILs in tumors larger than 0.022 g revealed significant differences of absolute cell numbers per 0.1 g tumor tissue for the following subsets CD8+IL-2+ (p = 0.01), CD8+IFN-γ+ (p = 0.01), CD8+TNF-α+ (p = 0.002), CD4+IL-2+ (p = 0.002), CD4+IFN-γ+ (p = 0.0006), CD4+TNF-α+ (p = 0.0009) in Nr2f6−/− mice.(B) qRT-PCR analysis of tumors revealed enhanced Ifng (p = 0.008) as well as Il2 expression (p = 0.052) in Nr2f6−/− mice. Results shown are derived from at least two independent experiments.(C) Dot plots and histograms of IFN-γ and IL-2 expressing CD8+ and CD4+ tumor-infiltrating cells gated on CD45+CD3+ T cells derived from Nr2f6+/+ and Nr2f6−/− mice. Numbers indicate percentage of positive gated cells. Results shown are derived from at least two independent experiments. Error bars represent SEM; data were analyzed via Student’s t test.(D) Tumor growth was monitored in Nr2f6−/− mice injected s.c. with 5 × 105 B16-OVA melanoma cells and administered with 0.5 mg of either an anti-mouse IFN-γ (n = 5) or an anti-mouse IL-2 (n = 3) neutralizing antibody or an corresponding IgG1 or IgG2a control every 3 days starting from day 1 of B16-OVA challenge. Comparison with relevant IgG control revealed disrupted tumor protection in Nr2f6−/− mice treated with anti-mouse IFN-γ neutralizing antibody (p = 0.006), but not anti-mouse IL-2 neutralizing antibody (p = 0.4). Summary graphs are the mean ± SEM, statistically analyzed by two-way ANOVA.
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fig4: Nr2f6 Expression Limits Cytokine Secretion of Tumor-Reactive T Cells(A) Cytokine secretion of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) TILs in tumors larger than 0.022 g revealed significant differences of absolute cell numbers per 0.1 g tumor tissue for the following subsets CD8+IL-2+ (p = 0.01), CD8+IFN-γ+ (p = 0.01), CD8+TNF-α+ (p = 0.002), CD4+IL-2+ (p = 0.002), CD4+IFN-γ+ (p = 0.0006), CD4+TNF-α+ (p = 0.0009) in Nr2f6−/− mice.(B) qRT-PCR analysis of tumors revealed enhanced Ifng (p = 0.008) as well as Il2 expression (p = 0.052) in Nr2f6−/− mice. Results shown are derived from at least two independent experiments.(C) Dot plots and histograms of IFN-γ and IL-2 expressing CD8+ and CD4+ tumor-infiltrating cells gated on CD45+CD3+ T cells derived from Nr2f6+/+ and Nr2f6−/− mice. Numbers indicate percentage of positive gated cells. Results shown are derived from at least two independent experiments. Error bars represent SEM; data were analyzed via Student’s t test.(D) Tumor growth was monitored in Nr2f6−/− mice injected s.c. with 5 × 105 B16-OVA melanoma cells and administered with 0.5 mg of either an anti-mouse IFN-γ (n = 5) or an anti-mouse IL-2 (n = 3) neutralizing antibody or an corresponding IgG1 or IgG2a control every 3 days starting from day 1 of B16-OVA challenge. Comparison with relevant IgG control revealed disrupted tumor protection in Nr2f6−/− mice treated with anti-mouse IFN-γ neutralizing antibody (p = 0.006), but not anti-mouse IL-2 neutralizing antibody (p = 0.4). Summary graphs are the mean ± SEM, statistically analyzed by two-way ANOVA.

Mentions: In tumor-bearing Nr2f6−/− mice, CD4+ and CD8+ tumor-infiltrating T cell subsets expressed significantly increased amounts of IL-2, IFN-γ, and TNF-α (Figures 4A–4C). Among the dLN-derived cells, CD4+ T cells producing IL-2 and IFN-γ as well as CD8+ T cells secreting IFN-γ were significantly more abundant in Nr2f6−/− mice (Figures S4A–S4C) as were activated CD4+ and CD8+ T cells (CD4+CD44hi; CD8+CD44hi) (Figure S4D). Notably, we could not detect any obvious alterations in tumor-infiltrating natural killer (NK) cells (DX5+), macrophages (CD11b+), or DC subsets (CD11c+CD11b+ and CD11c+CD8a+) in Nr2f6−/− compared to wild-type tumor-bearing mice (Figure S4E).


The Nuclear Orphan Receptor NR2F6 Is a Central Checkpoint for Cancer Immune Surveillance.

Hermann-Kleiter N, Klepsch V, Wallner S, Siegmund K, Klepsch S, Tuzlak S, Villunger A, Kaminski S, Pfeifhofer-Obermair C, Gruber T, Wolf D, Baier G - Cell Rep (2015)

Nr2f6 Expression Limits Cytokine Secretion of Tumor-Reactive T Cells(A) Cytokine secretion of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) TILs in tumors larger than 0.022 g revealed significant differences of absolute cell numbers per 0.1 g tumor tissue for the following subsets CD8+IL-2+ (p = 0.01), CD8+IFN-γ+ (p = 0.01), CD8+TNF-α+ (p = 0.002), CD4+IL-2+ (p = 0.002), CD4+IFN-γ+ (p = 0.0006), CD4+TNF-α+ (p = 0.0009) in Nr2f6−/− mice.(B) qRT-PCR analysis of tumors revealed enhanced Ifng (p = 0.008) as well as Il2 expression (p = 0.052) in Nr2f6−/− mice. Results shown are derived from at least two independent experiments.(C) Dot plots and histograms of IFN-γ and IL-2 expressing CD8+ and CD4+ tumor-infiltrating cells gated on CD45+CD3+ T cells derived from Nr2f6+/+ and Nr2f6−/− mice. Numbers indicate percentage of positive gated cells. Results shown are derived from at least two independent experiments. Error bars represent SEM; data were analyzed via Student’s t test.(D) Tumor growth was monitored in Nr2f6−/− mice injected s.c. with 5 × 105 B16-OVA melanoma cells and administered with 0.5 mg of either an anti-mouse IFN-γ (n = 5) or an anti-mouse IL-2 (n = 3) neutralizing antibody or an corresponding IgG1 or IgG2a control every 3 days starting from day 1 of B16-OVA challenge. Comparison with relevant IgG control revealed disrupted tumor protection in Nr2f6−/− mice treated with anti-mouse IFN-γ neutralizing antibody (p = 0.006), but not anti-mouse IL-2 neutralizing antibody (p = 0.4). Summary graphs are the mean ± SEM, statistically analyzed by two-way ANOVA.
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fig4: Nr2f6 Expression Limits Cytokine Secretion of Tumor-Reactive T Cells(A) Cytokine secretion of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) TILs in tumors larger than 0.022 g revealed significant differences of absolute cell numbers per 0.1 g tumor tissue for the following subsets CD8+IL-2+ (p = 0.01), CD8+IFN-γ+ (p = 0.01), CD8+TNF-α+ (p = 0.002), CD4+IL-2+ (p = 0.002), CD4+IFN-γ+ (p = 0.0006), CD4+TNF-α+ (p = 0.0009) in Nr2f6−/− mice.(B) qRT-PCR analysis of tumors revealed enhanced Ifng (p = 0.008) as well as Il2 expression (p = 0.052) in Nr2f6−/− mice. Results shown are derived from at least two independent experiments.(C) Dot plots and histograms of IFN-γ and IL-2 expressing CD8+ and CD4+ tumor-infiltrating cells gated on CD45+CD3+ T cells derived from Nr2f6+/+ and Nr2f6−/− mice. Numbers indicate percentage of positive gated cells. Results shown are derived from at least two independent experiments. Error bars represent SEM; data were analyzed via Student’s t test.(D) Tumor growth was monitored in Nr2f6−/− mice injected s.c. with 5 × 105 B16-OVA melanoma cells and administered with 0.5 mg of either an anti-mouse IFN-γ (n = 5) or an anti-mouse IL-2 (n = 3) neutralizing antibody or an corresponding IgG1 or IgG2a control every 3 days starting from day 1 of B16-OVA challenge. Comparison with relevant IgG control revealed disrupted tumor protection in Nr2f6−/− mice treated with anti-mouse IFN-γ neutralizing antibody (p = 0.006), but not anti-mouse IL-2 neutralizing antibody (p = 0.4). Summary graphs are the mean ± SEM, statistically analyzed by two-way ANOVA.
Mentions: In tumor-bearing Nr2f6−/− mice, CD4+ and CD8+ tumor-infiltrating T cell subsets expressed significantly increased amounts of IL-2, IFN-γ, and TNF-α (Figures 4A–4C). Among the dLN-derived cells, CD4+ T cells producing IL-2 and IFN-γ as well as CD8+ T cells secreting IFN-γ were significantly more abundant in Nr2f6−/− mice (Figures S4A–S4C) as were activated CD4+ and CD8+ T cells (CD4+CD44hi; CD8+CD44hi) (Figure S4D). Notably, we could not detect any obvious alterations in tumor-infiltrating natural killer (NK) cells (DX5+), macrophages (CD11b+), or DC subsets (CD11c+CD11b+ and CD11c+CD8a+) in Nr2f6−/− compared to wild-type tumor-bearing mice (Figure S4E).

Bottom Line: Nuclear receptor subfamily 2, group F, member 6 (NR2F6) is an orphan member of the nuclear receptor superfamily.Mechanistically, CD4(+) and CD8(+) T cell-intrinsic NR2F6 acts as a direct repressor of the NFAT/AP-1 complex on both the interleukin 2 and the interferon γ cytokine promoters, attenuating their transcriptional thresholds.Altogether, this defines NR2F6 as an intracellular immune checkpoint in effector T cells, governing the amplitude of anti-cancer immunity.

View Article: PubMed Central - PubMed

Affiliation: Translational Cell Genetics, Department for Pharmacology and Genetics, Medical University of Innsbruck, 6020 Innsbruck, Austria. Electronic address: natascha.kleiter@i-med.ac.at.

No MeSH data available.


Related in: MedlinePlus