Potential Antigens Involved In Delayed Xenograft Rejection in a GGTA1/CMAH DKO Pig-To-Monkey Model

When hyperacute rejection is avoided by deletion of Gal expression in the pig, delayed xenograft rejection (DXR) becomes a major immunologic barrier to successful xenotransplantation. This study was to investigate potential antigens involved in DXR. We isolated primary renal microvascular endothelial cells (RMEC) and aortic endothelial cells (AEC) from GGTA1/CMAH double-knockout (DKO) pig and immunized monkeys with both of these cells. After sensitization, monkey serum antibody binding and cytotoxicity to RMEC was significantlyhigher than to AEC,suggesting that RMEC are more immunogenic than AEC. Transcriptome sequencing of DKO pigs indicated that the expression of 1,500 genes was higher in RMEC than in AEC, while 896 genes was lower. The differentially-expressed genes were mainly related to cell and biological adhesion. Four kinds of cells were used for transcriptome sequencing, GGTA1/CMAH DKO pig AECs, GGTA1/CMAH DKO pig RMECs,cynomolgus monkey RMECs and human RMECs (HRGEC). Illumina Sequencing System (HiSeq2000, San Diego, CA, USA), following the manufacturer’s standard workflow. (This work was carried out by Novegene, Beijing, China).