Artificial antigen‐presenting cells (aAPCs) (NP‐SV95) are superior to mature dendritic cells (mDCs) ‐SV95, freshly prepared cytotoxic T lymphocytes (CTLs) are more potent than frozen CTLs, and the CTLs eradicate the target cells in an MHC I‐restricted manner. The cytolytic activities were compared between the CTLs that were induced with both aAPCs and mDCs. The aAPCs induced more effective and enhanced T‐cell responses in comparison to the mDCs [solid green line versus solid blue line, (a)]. The cytotoxicity of the SV95‐specific CTLs was significantly blocked by the addition of BB7.2 [broken red line versus solid red line; P values of 0·001, 0·002 and 0·007 for the E : T ratios of 50 : 1, 25 : 1 and 12·5 : 1, respectively, (b)]. In addition, the fresh CTLs had significantly greater activities than the frozen CTLs [solid green line versus solid red line; P values of 0·004, 0·007 and 0·006 for the E : T ratios of 50 : 1, 25 : 1 and 12·5 : 1, respectively, (c)]. Each cytotoxicity assay was independently repeated three times, and each condition was performed in triplicate. [Colour figure can be viewed at wileyonlinelibrary.com]