In this study, we first predicted four candidate epitopes from neuritin antigen by using HLA-A2.1-restricted epitope prediction algorithms based on long distance prediction systems SYFPEITHI and BIMAS. Secondly, peptide-binding assay was used to determine the affinity of every epitope with HLA-A2.1 and the results showed that neuritin13–21 had high affinity to HLA-A2.1, whereas neuritin121–129 and neuritin4–12 showed moderate affinity to the molecule. Thirdly, cytotoxic activity of CTLs was measured by ELISPOT and 51Cr release assay. The results demonstrated that neuritin13–21, neuritin121–129 and neuritin4–12 could elicit CTLs to lyse target cells in an HLA-A2.1-restricted manner. Lastly, we immunized the HLA-A*0201/Kb mice with various peptides and found neuritin13–21, neuritin121–129 and neuritin4–12 could also elicit CTLs to lyse neuritin and HLA-A2.1 positive target cells. These results suggested that the peptides had the potential of immunogenicity in vivo.