Figure 3
From: Monoclonal regulatory T cells provide insights into T cell suppression
(a) Representative flow cytometry plots show B3K506 Tregs from suppressive co-cultures stimulated with various B3K506 altered peptide ligands at a 1Tconv/4Treg ratio at 72h. Numbers in the plots depict % of cells in each quadrant. Bar graph shows mean number of B3K506 Tregs + /−SD from these cultures at 72 h, n = 5. (b) Representative histograms show CD25 expression (MFI) on CD4high Foxp3high B3K506 Tregs from suppressive co-cultures stimulated with various altered peptide ligands or no peptide at a 1Tconv/4Treg ratio at 72 h. (c) Representative histograms depict proliferation of CFSE labelled OT-II Tconvs form suppressive co-cultures described in A). Bar graph shows mean proliferation index + /−SD of OT-II Tconvs at 72 h, n = 4–8. (a–c) Statistical significance was calculated using a one-way ANOVA and subsequently a Bonferroni’s multiple comparisons test. NS = p > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (d) Curves show correlation between pMHC affinity (1/KD) for the B3K506 TCR and OT-II proliferation index from suppressive co-cultures described in A). APCs were pulsed with 10e-8M peptides (◽) or 10e-5M peptides (⚬). (e) Curve shows correlation between OT-II Tconv proliferation index and B3K506 Treg stimulus strength, which combines pMHC affinity and peptide concentration and is defined as [1/KD x peptide conc. (μM)] from suppressive co-cultures described in A). Curve is a nonlinear fit with variable slope (4 parameters). (f) Correlation between OT-II Tconv proliferation index and number of surviving Tregs from suppressive co-cultures described in A). Curve is a nonlinear fit with variable slope (4 parameters). (g) Correlation between OT-II Tconv proliferation index and CD25 expression (MFI) on surviving Tregs from suppressive co-cultures described in A). Curve is a nonlinear fit with variable slope (4 parameters).
