Figure 5. Concurrent Agonistic Anti-CD40 Treatment Allows Costimulation Blockade-Treated Alloreactive T Cell Populations to Undergo Avidity Maturation and Expansion.

(A) Normalized MFI of Foxp3⁻ 2W:I-A^b and OVA:K^b multimer-reactive splenic T cells from naive, rejected, and tolerant mice. In addition, a group of transplanted mice received a single concurrent dose of agonistic anti-CD40 on day 0 at the same time as the DST and anti-CD154 used for tolerance induction. (B) Numbers of splenic T conventional cells binding 2W:I-A^b and OVA:K^b. All transplanted mice were analyzed 8–9 days post-transplantation. (C and D) Normalized multimer and TCRβ (D) MFI of Foxp3⁻ 2W:I-A^b and OVA:K^b multimer-reactive graft-infiltrating T cells. (E) Numbers of graft-infiltrating T conventional cells binding 2W:I-A^b and OVA:K^b. Data points in naive, rejected, and tolerant groups were also presented in Figures S1 and 1C. Data are pooled from 2–5 independent experiments, including the Tol + anti-CD40 data that are pooled from two independent experiments, with n = 6 mice total per group. Box-and-whisker plots with Tukey whiskers are superimposed with the individual data points. Data were analyzed 8–9 days post-transplantation by one-way ANOVA with Bonferroni correction for multiple testing (A, C, and D), Kruskal-Wallis with Dunn’s test (E), or Student’s t test (B). ^**p < 0.01, ^***p < 0.001. NS, not significant. See also Figures S4 and S5.