Figure 1. Graft-Specific T Cells from Tolerant Mice Bind Less pMHC Multimer Than Cells from Rejecting Mice.

(A) Representative flow cytometry plots of splenic Foxp3⁺ and Foxp3⁻ T cell populations reactive to 2W:I-A^b and OVA:K^b isolated from naive untransplanted B6 mice, mice transplanted 8–9 days prior with a BALB/c × B6 F1 2W-mOVA or BALB/c-OVA heart and left untreated to undergo acute rejection (Rej), or mice transplanted and tolerized with anti-CD154 and DST (Tol). (B–E) Histograms and relative 2W:I-A^b and OVA:K^b multimer mean fluorescence intensity (MFI) reflecting multimer staining (B and C), TCRβ (B and C), and CD8α (D and E) levels of endogenous alloreactive T cells over time in naive, acute rejection, and tolerant recipients. Values were normalized, with the average of the MFI for the naive mice in each experiment set to 1. Data are pooled from 2–19 independent experiments. Box-and-whisker plots with Tukey whiskers are superimposed with the individual data points. Numbers of mice per group are listed below each panel. Data were analyzed by one-way ANOVA with Bonferroni correction for multiple testing. ^**p < 0.01, ^***p < 0.001 (for comparisons of naive, day 8–9 Rej, and day 8–9 Tol), ^###p < 0.001 (for comparisons of naive, day 60+ Rej, and day 60+ Tol). NS, not significant. See also Figures S1 and S2.