Objective:　To　predict　and　identify　the　leukemia-associated　antigen　MLAA-34　HLA-A2　restricted　cytotoxic　T　lymphocyte　(CTL)　epitopes　by　bio-informatic　methods　so　as　to　provide　the　probability　of　MLAA-34-based　target　for　leukemia　immunotherapy.　Methods:　MLAA-34　specific　HLA-A2　restricted　CTL　epitopes　were　predicted　by　computer　supermotif　algorithm　combined　with　quantitative　motif　algorithm.　Candidate　epitopes　were　verified　when　their　scores　were　relatively　high　and　in　the　top　10,　and　then　artificially　synthesized.　Affinity　of　the　candidate　epitope　was　examined　by　HLA-A2　binding　assay　combined　with　flow　cytometry　applying　T2　cells　(shown　as　fluorescence　index,　FI).　Four　selected　candidates　with　higher　affinity　were　detected　by　lactate　dehydrogenase　(LDH)　release　assay　kit　to　determine　their　ability　to　induce　the　generation　of　specific　CTLs　in　vitro.　Results:　Among　the　top　10　candidate　CTL　epitope　peptide　derived　from　MLAA-34,　MLAA2　(ILLKNQPKL),　MLAA3　(LLTRHKVLV),　MLAA5　(LLVTLIADL)　and　MLAA9　(YLIKQIRDL)　had　a　higher　affinity　of　HLA-A2;　the　FI　values　were　1.65,　1.73,　1.82　and　1.05,　respectively.　These　epitope　peptides　could　be　candidates　for　further　analysis.　The　analysis　of　the　CTLs　cytotoxicity　showed　that　the　peptide　MLAA5　(LLVTLIADL)　could　effectively　induce　specific　CTLs　in　vitro,　and　the　CTLs　could　lyse　not　only　the　human　leukemia　cell　line　THP-1　but　also　the　cell　line　T2　pulsed　with　MLAA3　(LLTRHKVLV).　Conclusion:　MLAA5　(LLVTLIADL)　is　a　CTL　epitope　of　the　leukemia-associated　antigen　MLAA-34　presented　by　HLA-A2.　This　study　provides　an　experimental　basis　for　design　and　preparation　of　therapeutic　vaccines　based　on　MLAA-34.