Polymorphonuclear neutrophils (PMNs), traditionally considered effector cells in the inflammatory response, have recently been regarded as potential regulators of the immune response. In the present study we investigate whether PMNs are efficient antigen‐presenting cells for reactivation of memory cytotoxic T lymphocytes (CTLs). PMNs were pulsed with synthetic peptides derived from Epstein‐Barr virus (EBV) antigens. We have used the IVTDFSVIK (IVT) peptide derived from the Epstein‐Barr virus—encoded nuclear antigen 4 protein, corresponding to the immunodominant epitope of HLA‐A11–restricted CTL responses, and the CLGGLLTMV (CLG) peptide derived from the latent membrane protein 2 antigen, representing a subdominant epitope of HLA‐A2–restricted CTL responses. The data indicate that peptide‐pulsed PMNs selectively activate specific CTL responses to both immunodominant and subdominant epitopes. The efficiency of CTL induction by PMNs was comparable to that observed with the conventional method of EBV‐specific CTL reactivation with the autologous lymphoblastoid cell line, as well as with peptide‐pulsed monocyte‐enriched adherent cells. On the contrary, unactivated peptide‐pulsed lymphocytes failed to induce an epitope‐specific CTL response. These results demonstrate that PMNs efficiently present antigens to memory virus‐specific CTLs and suggest that they may have a role as antigen‐presenting cells. J. Leukoc. Biol. 60: 207–213; 1996.