DIRECT RECOGNITION OF SLA-LIKE AND HLA-LIKE CLASS-II ANTIGENS ON PORCINE ENDOTHELIUM BY HUMAN T-CELLS RESULTS IN T-CELL ACTIVATION AND RELEASES OF INTERLEUKIN-2

To investigate whether human T cells can directly recognize pig xenoantigens, highly purified human CD4(+) and CD8(+) T cells were incubated with pig aortic endothelial cells (PAEC). The response was measured by [H-3]thymidine uptake and release of bioactive interleukin-2. A detailed examination of MHC expression by cultured PAEC and tissue sections of porcine aorta and heart showed porcine endothelial cells (EC) to be constitutively positive for SLA class II and antigens that crossreact with HLA class II molecules. Low level expression of B7 receptors was detected by binding of both human and mouse CTLA-4-Ig to untreated PAEC, which was enhanced significantly by treatment with recombinant porcine interferon-gamma. Human T cells, purified by positive selection and residual DR(+) cells removed by lymphocytolysis, were shown to be functionally free of monocytes. Untreated PAEC elicited strong proliferation by human CD4(+) T cells: CD8(+) T cells also proliferated, but more weakly. This response was inhibited by CTLA-4-Ig. Blocking studies were performed with mAbs that bind to PAEC and not human EC (MSA3, TH16B), an mAb that binds to human and porcine EC (DA6.231), and L243, which binds to human and not porcine EC. The proliferative response of CD4(+) T cells to PAEC was inhibited significantly by mAbs against swine and human determinants. In contrast, the response of CD4(+) T cells to human EC was inhibited only by mAbs against human determinants. Experiments that directly compared the CD4(+) and CD8(+) T cell responses to PAEC and the human EC Line EAhy.926, both with and without prior treatment with species-specific interferon gamma, demonstrated greater proliferation and 5-10 times more interleukin-2 in response to pig EC than to human EC.