Recognition of HLA-A*0201-matched KDR+ aECs by K288.E5. (A) HLA-A2 and KDR phenotypes are summarized for each HUVEC and HDMEC primary culture. KDR protein in 30 μg cell lysate was assessed by Western blot. (B) Lysis of individual aEC cultures by K288.E5 at different E/T ratios. Cytolysis was determined in a 16-hour LDH release assay. (C) T-cell recognition of HLA-mismatched aECs on transfection with the HLA-A*0201 gene. Transfection and T-cell assays were performed as described in Figure 2. All experiments were performed 3 times, with similar results. (D) Cold-target inhibition assays containing unlabeled targets (1 × 105/well), 51Cr-labeled HUVEC2 cells (5 × 103/well), and K288.E5 at indicated E/T ratios. Cytotoxic activities were determined in a 6-hour 51Cr release assay. Data shown are representative of 2 independent experiments.
