Blockade of the PD‐1/PD‐1L pathway reverses the protective effect of anti‐CD40L therapy in a rat to mouse concordant islet xenotransplantation model

  Background:  We have previously demonstrated that costimulatory blockade with anti‐CD40L monoclonal antibody (mAb) prolongs the survival of non‐vascularized concordant rat to mouse islet xenografts. Here, we examine whether signaling through the PD‐1/PD‐1L pathway is required for the anti‐CD40L therapy to prolong concordant islet graft survival using a novel anti‐murine PD‐1 mAb (clone 4F10). Methods:  C57BL/6 mice received a cellular concordant islet xenograft under the left kidney capsule and four experimental groups were prepared. Group I: untreated control; group II: recipient mice were treated with three doses of 0.5 mg of anti‐CD40L mAb (clone MR1) on days 0, 2 and 4; group III: mice were treated with 0.5 mg of anti‐PD‐1 (CD279) mAb (clone 4F10) every other day for 8 days; and finally group IV: mice received the combined treatment that consisted of anti‐CD40L plus anti‐PD‐1 mAb. Results:  Concordant islet xenografts transplanted in control untreated mice showed a median survival time (MST) of 17 ± 7.43 days, whereas anti‐CD40L treatment led to a significant prolongation of graft survival (MST: 154 ± 65.56, P < 0.0001). The administration of anti‐PD‐1 alone significantly accelerated graft rejection compared to non‐treated controls (MST: 10 ± 2.24 vs. MST: 17 ± 7.43, P < 0.0004). Remarkably, the combined administration of anti‐CD40L and anti‐PD‐1 reversed the protective effect obtained with anti‐CD40L alone (anti‐CD40L, MST: 154 ± 65.56 vs. anti‐CD40L plus anti‐PD‐1, MST: 10 ± 7.72, P < 0.0002). Conclusion:  Overall, our data indicate that the PD‐1/PD‐1L pathway is required for the achievement of prolonged graft survival in anti‐CD40L‐treated mice in a setting of rat to mouse concordant islet xenotransplantation.