Results: 3

1.
Fig. 3.

Fig. 3. From: CD4+CD25+Foxp3+ regulatory T cell formation requires more specific recognition of a self-peptide than thymocyte deletion.

Agonist and partial agonist self-peptides activate distinct signaling pathways during 6.5hiCD4SP thymocyte development. (A) Representative histograms showing intracellular Nur77 expression by 6.5hiCD4SPCD24hi thymocytes from TS1 × HA28 (red) (n = 4), TS1 × PevHA (green) (n = 4), and TS1 × Pev(SW)HA (blue) (n = 4) mice are shown relative to TS1 (filled) (n = 4) mice. Mean fluorescent intensities (MFIs) of Nur77 staining are shown ± SEM. **P ≤ 0.05, ***P ≤ 0.01. (B) Representative (n = 4) dot plots showing Foxp3 versus CD25 expression by 6.5hiCD4SPCD24hi (Upper) and 6.5hiCD4SPCD24low (Lower) thymocytes from TS1, TS1 × HA28, TS1 × PevHA, and TS1 × Pev(SW)HA mice. Mean percentages of cells in the indicated quadrants are shown. (C) Graph shows mean percentages ± SEM of 6.5hiCD4SP thymocytes that are Foxp3+ (open bars) or Nur77+ (filled bars) in TS1 (n = 4), TS1 × HA28 (n = 4), TS1 × PevHA (n = 4), and TS1 × Pev(SW)HA (n = 4) mice. **P ≤ 0.05, ***P ≤ 0.01. All data are representative of at least three independent experiments.

Cristina Cozzo Picca, et al. Proc Natl Acad Sci U S A. 2011 September 6;108(36):14890-14895.
2.
Fig. 1.

Fig. 1. From: CD4+CD25+Foxp3+ regulatory T cell formation requires more specific recognition of a self-peptide than thymocyte deletion.

The partial agonist S1(SW) peptide induces 6.5hi thymocyte deletion but not Treg cell formation. (A) Proliferation of 6.5hiCD4+ T cells from TS1 mice incubated with graded doses of S1 peptide (Left) or with graded numbers of HAU of PR8 virus (circles) or SW virus (triangles) (Right) shown as means ± SEM. n = 3. (B) Representative histograms show 6.5 and Foxp3 expression by CD4SP thymocytes from TS1 × HA28 (n = 5), TS1 × PevHA (n = 6), and TS1 × Pev(SW)HA (n = 6) mice (black lines) overlaying thymocytes from TS1 (n = 11) mice (filled), with mean percentages in gate indicated. Graphs show numbers of 6.5hiCD4SP and 6.5hiCD4SPFoxp3+ thymocytes, with bars denoting means and individual values shown. **P ≤ 0.05, ***P ≤ 0.01. (C) Same as in B except showing 6.5, CD25, and Foxp3 expression by CD4+ LN cells. (D) Representative histograms show Foxp3 expression on 6.5+CD4SP thymocytes from chimeras of Pev(SW)HA (n = 6) or PevHA (n = 6) mice (black lines) overlaying filled histograms from BALB/c (n = 6) mice (shaded) reconstituted with BM from TS1.RAG−/− mice. Mean percentages in indicated gates are shown. Graph shows numbers of thymocytes with bars denoting means and individual values shown. **P ≤ 0.05. All data are representative of at least three independent experiments.

Cristina Cozzo Picca, et al. Proc Natl Acad Sci U S A. 2011 September 6;108(36):14890-14895.
3.
Fig. 2.

Fig. 2. From: CD4+CD25+Foxp3+ regulatory T cell formation requires more specific recognition of a self-peptide than thymocyte deletion.

The TS1(SW) TCR undergoes Treg cell formation in response to agonist S1(SW) peptide. (A) Vα8.3+CD4+ T cells from TS1(SW) mice incubated with graded doses of S1(SW) peptide (Left) or with graded numbers of HAU of PR8 virus (circles) or SW virus (triangles) (Right), shown as means ± SEM. n = 3. (B) Representative histograms showing Vα8.3 and Foxp3 expression by CD4SP thymocytes from TS1(SW) × PevHA (n = 5) and TS1(SW) × Pev(SW)HA (n = 5) mice (black lines) overlaying thymocytes from TS1(SW) (n = 12) mice (filled), with mean percentages in gate indicated. Graphs show numbers of Vα8.3+CD4SP and Vα8.3+CD4SPFoxp3+ thymocytes, with bars denoting means and individual values shown. (C) Same as in B except showing Vα8.3, CD25 ,and Foxp3 expression by CD4+ LN cells. ***P ≤ 0.01. (D) Representative histograms showing Vα8.3 and Foxp3 expression by CD4SP thymocytes in chimeric Pev(SW)HA (n = 6) or PevHA (n = 6) mice (black lines) overlaying filled histograms from BALB/c (n = 6) mice (shaded) reconstituted with BM from TS1(SW).RAG−/− mice. Mean percentages in indicated gates are shown. Graph shows numbers of thymocytes with bars denoting means and individual values shown. **P ≤ 0.05. (E) Representative histograms showing Vα8.3 and Foxp3 expression by CD4SP thymocytes in chimeric BALB/c (filled) or Pev(SW)HA (black) mice reconstituted with 100% BM from TS1(SW) mice (n = 4) (Top), 10% BM from TS1(SW) mice plus 90% BM from BALB/c mice (n = 4) (Middle), or 100% BM from BALB/c mice (n = 4) (Bottom). Mean percentages in indicated gates in BALB/c (gray) and Pev(SW)HA (black) recipients are shown. All data are representative of at least three independent experiments.

Cristina Cozzo Picca, et al. Proc Natl Acad Sci U S A. 2011 September 6;108(36):14890-14895.

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