T cell receptor repertoire profiling of murine skin transplantation in context of PD-1+ cells Ablation therapy

The purpose of this study was to evaluate the influence of ablating PD-1+ cells on alloreactive T cell repertoires. We established the murine skin transplantation model and obtained the draining lymph nodes (dLNs) T cells from recipients for TCR repertoires sequencing. PD-1DTR mice were generated via knocking IRES-DTR-P2A-EGFP-P2A-firefly Luciferase into 3’ UTR of the Pdcd1 locus with EGE (CRISPR/Cas9) system from Biocytogen (Beijing, China). 4 to 6-week-old male mice were performed with thymectomy and recovered for one month before transplantation. The murine skin transplantation model was established as previously described. In the ISO group, the dorsal sides of C57BL/6 (B6; H-2b) ears were transplanted onto the dorsum of PD-1DTR recipients. While in the PBS group and DT group, BALB/c (H-2d) ears were transplanted onto the dorsum of PD-1DTR recipients, with PBS or DT (10μg/kg) treatment. At day 9 post transplantation, dLNs of recipients were obtained with Pan T Cell Isolation Kit for TCR repertoires sequencing. Each group has 3 replicates. Each replicate was generated from isolated T cells pooled from 3 mice.