TNFα/TNFR2 disruption and its effect on GVHD do not depend on the antigen specificity of therapeutic Tregs. (A-B) [B6 x C3H]F1 female mice underwent TBI followed by transplantation with (1) B6 BM cells plus 2 × 10⁶ T cells or (2) B6 BM cells plus 2 × 10⁶ T cells supplemented with 2 × 10⁶ rs-Tregs or (3) BM cells plus 2 × 10⁶ T cells collected from TNFα-deficient mice supplemented with 2 × 10⁶ rs-Tregs. HY peptide was administered at day 0, 1, 3, and 6, and mice were treated or not with anti-TNFR2 administered at day 0, 2, and 4. The resulting survival (A) and clinical score (B) data were compared among the 3 groups of mice. Mice were euthanized in case of weight loss >30% of initial weight or maximal clinical grade (ie, 5/5). Kaplan-Meier survival curves were compared using log-rank test. For analysis of GVHD clinical grading curves, AUC was calculated for each mouse, and then 1-way ANOVA with post hoc analysis was performed. *P < .05; **P < .01; ***P < .001.