PD-L1 is required for human endometrial regenerative cells-associated attenuation of experimental colitis in mice

Endometrial regenerative cells (ERCs) are easily isolated from menstrual blood, and can be cultured in large amounts. Although, ERCs can ameliorate DSS-induced colitis in mice, the molecular mechanism underlying ERCs-mediated immunosuppression is unclear. This study was aimed to assess the function of PD-L1 expressed on ERCs in colitis attenuation. ERCs with and without anti-PD-L1 mAb-pretreatment were administered to mice by injection at 2, 5 and 8 days after colitis induction by DSS treatment. Blood, spleen and colon samples were obtained 15 days post-DSS-induction. Then, clinicopathological alterations, cytokine levels, immune cell types and cell tracking were assessed. ERCs or ERCs preincubated with anti-PD-L1 antibody were co-cultured with splenocytes, whose phenotypes was analyzed by flow cytometry. We found that PD-L1 on ERCs was upregulated by IFN-γ stimulation. The transplanted PKH26-labeled ERCs were engrafted to the lung, liver, spleen and injured colon. Interestingly, ERC-based therapy markedly attenuated mouse colitis, but blockade of PD-L1 on ERCs with a specific monoclonal antibody conferred severe colitis to the animals. These effects of PD-L1 inhibition on colitis were associated with reduced amounts of pro-inflammatory cytokines and infiltrated immune cells, including CD3⁺CD4⁺ T lymphocytes, CD3⁺CD8⁺ T lymphocytes, CD11c⁺MHC-II⁺ Dendritic cells and F4/80⁺ macrophages, both in vivo and in vitro, as well as with elevated levels of anti-inflammatory cytokines and regulatory immune cells, including CD4⁺CD25⁺Foxp3⁺ Tregs and F4/80⁺CD206⁺ macrophages. These findings demonstrated that ERCs-based treatment promotes immune tolerance in mouse colitis, in association with PD-L1, thus indicating that PD-L1 modulates immunosuppression by ERCs.