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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

Enhanced Numbers of Tumor-Infiltrating T Cells in Nr2f6-Deficient Mice Bearing B16-OVA Transplantable Melanomas(A) Gross examination of B16-OVA melanomas from Nr2f6+/+ (top) and Nr2f6−/− (bottom) mice 14 days after subcutaneous injection of 1 × 105 tumor cells.(B) Tabulation of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells on day 14. Significantly increased absolute cell numbers of CD45+ (p = 0.01), CD3+ (p = 0.05), CD8+ (p = 0.007), and CD4+ (p = 0.015) per 0.1 g of tumor tissue were detected in Nr2f6−/− mice.(C) Dot plots of B16-OVA tumor-infiltrating CD45+ and CD3+ cells derived from Nr2f6+/+ and Nr2f6−/− tumor-bearing mice (for detailed gating strategy, see Figure S3).(D) Immune cell characterization of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells per 0.1 g of tumor tissue, gated on CD45+CD3+ positive cells did reveal increased absolute cell numbers of CD8+CD25+ (p = 0.003); CD8+PD-1+ (p = 0.003); CD4+CD25+ (p = 0.008); CD4+PD-1+ (p = 0.001); and CD4+CD25+Foxp3+ Treg cells (p = 0.007).(E) Dot plots of B16-OVA tumor-infiltrating CD8+PD-1+ cells derived from Nr2f6+/+ and Nr2f6−/− mice.(F) Significantly increased absolute cell numbers of CD11b+Gr-1+ myeloid derived suppressor cells (p = 0.02), and a trend for tumor associated macrophages (p = 0.06) (CD11b+F/80+) could be detected in tumors of Nr2f6−/− (n = 7) mice.
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fig3: Enhanced Numbers of Tumor-Infiltrating T Cells in Nr2f6-Deficient Mice Bearing B16-OVA Transplantable Melanomas(A) Gross examination of B16-OVA melanomas from Nr2f6+/+ (top) and Nr2f6−/− (bottom) mice 14 days after subcutaneous injection of 1 × 105 tumor cells.(B) Tabulation of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells on day 14. Significantly increased absolute cell numbers of CD45+ (p = 0.01), CD3+ (p = 0.05), CD8+ (p = 0.007), and CD4+ (p = 0.015) per 0.1 g of tumor tissue were detected in Nr2f6−/− mice.(C) Dot plots of B16-OVA tumor-infiltrating CD45+ and CD3+ cells derived from Nr2f6+/+ and Nr2f6−/− tumor-bearing mice (for detailed gating strategy, see Figure S3).(D) Immune cell characterization of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells per 0.1 g of tumor tissue, gated on CD45+CD3+ positive cells did reveal increased absolute cell numbers of CD8+CD25+ (p = 0.003); CD8+PD-1+ (p = 0.003); CD4+CD25+ (p = 0.008); CD4+PD-1+ (p = 0.001); and CD4+CD25+Foxp3+ Treg cells (p = 0.007).(E) Dot plots of B16-OVA tumor-infiltrating CD8+PD-1+ cells derived from Nr2f6+/+ and Nr2f6−/− mice.(F) Significantly increased absolute cell numbers of CD11b+Gr-1+ myeloid derived suppressor cells (p = 0.02), and a trend for tumor associated macrophages (p = 0.06) (CD11b+F/80+) could be detected in tumors of Nr2f6−/− (n = 7) mice.

Mentions: As next step the B16-OVA subcutaneous (s.c.) tumor model was used for an extensive analysis of tumor-infiltrating immune cells at day 14 after tumor cell injection (Figure 3A), employing a stratified CD45+/CD3+/CD4+ or CD8+ gating strategy (as outlined in Figures S3A and S3B). As in the autochthonous model of prostate cancer, B16-OVA tumors grown in Nr2f6−/− mice exhibited significantly increased numbers of tumor-infiltrating CD45+CD3+CD4+ and CD45+CD3+CD8+ T cells, calculated at the level of total numbers on a weight (i.e., gram) basis when compared to wild-type mice (Figures 3B and 3C). This particular phenotype of Nr2f6−/− mice was independent of the B16-OVA tumor size (Figure S3C). Analysis in tumors with comparable sizes by immunohistochemistry confirmed the markedly enhanced intratumoral CD4+ and CD8+ T cell numbers in Nr2f6−/− mice (Figures S3D and S3E), well reflecting relative levels of tumor-infiltrating lymphocytes (TILs) (i.e., percentage of CD45+, CD45+CD3+, CD45+CD8+, or CD45+CD4+cells; Figure S3F). Pronounced CD8+ and CD4+ T cell effector functions in Nr2f6−/− mice were found to be associated with markedly higher expression of CD25 and PD-1 on both T cell subsets when compared to wild-type animals (Figures 3D and 3E).


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)

Enhanced Numbers of Tumor-Infiltrating T Cells in Nr2f6-Deficient Mice Bearing B16-OVA Transplantable Melanomas(A) Gross examination of B16-OVA melanomas from Nr2f6+/+ (top) and Nr2f6−/− (bottom) mice 14 days after subcutaneous injection of 1 × 105 tumor cells.(B) Tabulation of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells on day 14. Significantly increased absolute cell numbers of CD45+ (p = 0.01), CD3+ (p = 0.05), CD8+ (p = 0.007), and CD4+ (p = 0.015) per 0.1 g of tumor tissue were detected in Nr2f6−/− mice.(C) Dot plots of B16-OVA tumor-infiltrating CD45+ and CD3+ cells derived from Nr2f6+/+ and Nr2f6−/− tumor-bearing mice (for detailed gating strategy, see Figure S3).(D) Immune cell characterization of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells per 0.1 g of tumor tissue, gated on CD45+CD3+ positive cells did reveal increased absolute cell numbers of CD8+CD25+ (p = 0.003); CD8+PD-1+ (p = 0.003); CD4+CD25+ (p = 0.008); CD4+PD-1+ (p = 0.001); and CD4+CD25+Foxp3+ Treg cells (p = 0.007).(E) Dot plots of B16-OVA tumor-infiltrating CD8+PD-1+ cells derived from Nr2f6+/+ and Nr2f6−/− mice.(F) Significantly increased absolute cell numbers of CD11b+Gr-1+ myeloid derived suppressor cells (p = 0.02), and a trend for tumor associated macrophages (p = 0.06) (CD11b+F/80+) could be detected in tumors of Nr2f6−/− (n = 7) mice.
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fig3: Enhanced Numbers of Tumor-Infiltrating T Cells in Nr2f6-Deficient Mice Bearing B16-OVA Transplantable Melanomas(A) Gross examination of B16-OVA melanomas from Nr2f6+/+ (top) and Nr2f6−/− (bottom) mice 14 days after subcutaneous injection of 1 × 105 tumor cells.(B) Tabulation of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells on day 14. Significantly increased absolute cell numbers of CD45+ (p = 0.01), CD3+ (p = 0.05), CD8+ (p = 0.007), and CD4+ (p = 0.015) per 0.1 g of tumor tissue were detected in Nr2f6−/− mice.(C) Dot plots of B16-OVA tumor-infiltrating CD45+ and CD3+ cells derived from Nr2f6+/+ and Nr2f6−/− tumor-bearing mice (for detailed gating strategy, see Figure S3).(D) Immune cell characterization of Nr2f6+/+ (n = 6) and Nr2f6−/− (n = 7) B16-OVA tumor-infiltrating cells per 0.1 g of tumor tissue, gated on CD45+CD3+ positive cells did reveal increased absolute cell numbers of CD8+CD25+ (p = 0.003); CD8+PD-1+ (p = 0.003); CD4+CD25+ (p = 0.008); CD4+PD-1+ (p = 0.001); and CD4+CD25+Foxp3+ Treg cells (p = 0.007).(E) Dot plots of B16-OVA tumor-infiltrating CD8+PD-1+ cells derived from Nr2f6+/+ and Nr2f6−/− mice.(F) Significantly increased absolute cell numbers of CD11b+Gr-1+ myeloid derived suppressor cells (p = 0.02), and a trend for tumor associated macrophages (p = 0.06) (CD11b+F/80+) could be detected in tumors of Nr2f6−/− (n = 7) mice.
Mentions: As next step the B16-OVA subcutaneous (s.c.) tumor model was used for an extensive analysis of tumor-infiltrating immune cells at day 14 after tumor cell injection (Figure 3A), employing a stratified CD45+/CD3+/CD4+ or CD8+ gating strategy (as outlined in Figures S3A and S3B). As in the autochthonous model of prostate cancer, B16-OVA tumors grown in Nr2f6−/− mice exhibited significantly increased numbers of tumor-infiltrating CD45+CD3+CD4+ and CD45+CD3+CD8+ T cells, calculated at the level of total numbers on a weight (i.e., gram) basis when compared to wild-type mice (Figures 3B and 3C). This particular phenotype of Nr2f6−/− mice was independent of the B16-OVA tumor size (Figure S3C). Analysis in tumors with comparable sizes by immunohistochemistry confirmed the markedly enhanced intratumoral CD4+ and CD8+ T cell numbers in Nr2f6−/− mice (Figures S3D and S3E), well reflecting relative levels of tumor-infiltrating lymphocytes (TILs) (i.e., percentage of CD45+, CD45+CD3+, CD45+CD8+, or CD45+CD4+cells; Figure S3F). Pronounced CD8+ and CD4+ T cell effector functions in Nr2f6−/− mice were found to be associated with markedly higher expression of CD25 and PD-1 on both T cell subsets when compared to wild-type animals (Figures 3D and 3E).

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