Transcriptome Sequencing Reveals Molecular Signatures of Antigen-encountering T cells in Adoptive transfer Murine Heart Transplantation Model

The purpose of this study was to identify the molecular signatures of antigen-encountering T cells, we established the adoptive transfer murine heterotopic heart transplantation model and isolated TEa cells from spleens for RNA sequencing. Overall design: B6.Cg-Tg(Tcra,Tcrb)3Ayr/J(TEa TCR transgenic mice) and B6.SJL-Ptprca Pepcb/BoyJ (B6 CD45.1) mice were purchased from Jackson Laboratories (Bar Harbor, ME). TEa cells were isolated from spleen of TEa mice via CD4+ T Cell Isolation Kit (70901-100; BEAVER). And the purity was checked with FCM (>90%). The B6 CD45.1+ recipients were intravenously transferred with 5 million CD45.2+ TEa cells one day before heart transplantation. The murine heterotopic heart transplantation model was established as previously described. In Allograft group, BALB/c (H-2d) hearts were transplanted into CD45.1+ recipients. While in Isograft group, C57BL/6 (B6; H-2b) hearts were used as the grafts. On day 6, the TEa cells were sorted from the recipients' spleens, and differentially expressed genes between the allograft and isograft groups were determined by RNA-seq. Each group has 3 replicates. Each replicate was generated from sorted TEa cells pooled from 3 mice.