Wild‐type (WT) sequence p53 peptides are attractive candidates for broadly applicable cancer vaccines. The aim of this study was to evaluate the potential of a WT p53‐based immunotherapeutic approach for patients with hepatocellular carcinoma (HCC). Circulating CD8⁺ T cells specific for WT p53_149–157 and WT p53_264–272 HLA‐A*0201 restricted epitopes were directly identified in the peripheral blood by the use of peptide/HLA‐A2.1 tetramers in 24 HCC patients. Cytotoxic T lymphocyte (CTL) activity after WT p53 peptide‐specific stimulation was assessed by analysis of granzyme B and interferon‐gamma mRNA transcription, using a quantitative real‐time polymerase chain reaction assay. Tumor immunophenotyping was performed to evaluate the p53 status, the expression of major histocompatibility complex (MHC) and costimulatory molecules in freshly isolated tumor cells. HCC patients exhibited significantly higher frequencies of WT p53‐specific memory CD8⁺ T cells and stronger WT p53‐specific CTL activity, when compared with healthy controls. Increased frequencies of p53‐specific CD8⁺ T cells and their activity correlated with selective HLA‐A2 allele loss and reduced costimulatory molecule expression of tumor cells. Moreover, augmented numbers of p53‐specific T cells coincided with high MHC class II expression in tumor cells but were inversely related to the T status of the tumor node metastasis staging system. Our results indicate the existence of natural immunosurveillance and tumor immune evasion, involving a T cell response against WT p53 tumor antigen in patients with HCC. These findings may have important implications for the future development of cancer vaccines. © 2006 Wiley‐Liss, Inc.