The use of HLA A2,1/p53 peptide tetramers to visualize the impact of self tolerance on the TCR repertoire

p53 is an attractive target for cancer immunotherapy since it is overexpressed in half of all tumors. However, it is also expressed in normal lymphoid tissue, and self tolerance leaves a p53-specific repertoire purged of high avidity CTL, To better understand the mechanism of tolerance and the basis for such low avidity interaction, p53-specific CTL from p53 deficient (p53(-)) and sufficient (p53(+)) A2.1/K-b transgenic mice were compared with respect to their ability to bind HLA-A2.1 tetramers containing cognate murine p53 peptide Ag, p53 261-269, Since the murine CD8 molecule cannot interact with human KLA-A2.1, this tests the ability of the TCR to bind the A2/1.peptide complex: tetramer, CTL from p53- mice demonstrated strong binding of such A2.1/p53 261-269 tetramers; however, the CTL from tolerant p53+ mice were devoid of tetramer-binding CD8(+) T cells. Examination of TCR expression at the clonal level revealed that CTL from p53(+) and p53- mice each expressed comparable levels of the p53-specific TCR, These results indicate that normal expression of p53 promotes elimination of T cells expressing TCRs with sufficient affinity to achieve stable binding of the A2.1/p53 261-269 tetramers.