Flow cytometric analysis and cellular assays were used to determine constitutive and induced expression of class I HLA antigens and class II antigens encoded by the HLA-DR, -DQ and -DP subregions on cultured human arterial endothelial cells (HAEC) derived from transplant donors. Class I but no or minimal quantities of class II HLA antigens were found on HAEC. Prior incubation of HAEC with gamma-IFN increased class I HLA antigen expression and induced class II HLA antigen expression on HAEC. The induced expression of HLA-DQ was lower than that of HLA-DR, but both were significantly reduced in comparison to the frequency of these antigens on EBV transformed lymphoblastoid cell lines derived from the same donor. In addition, supernatants from class I and class II alloreactive clones were shown to induce class II antigen expression on HAEC. By PLT analysis, it was shown that these antigens are functionally capable of generating a lymphocyte response. In this regard, HAEC have proved to be a helpful tool in designing in vitro lymphocyte-endothelial cell studies.