Methods and compositions for producing target cell reactive lymphocytes, e.g., cytolytic T-lymphocytes, in a subject are provided. In practicing the subject methods, a lymphocyte population is contacted with an effective amount of a target cell peptide mixture of active and inactive peptides to produce lymphocytes reactive, e.g., cytolytic, for the target cell. Also provided are compositions, kits, and systems for practicing the subject methods. The subject invention finds use in a variety of different applications, including therapeutic applications.