Expression of reporter genes in muscle cells has been achieved by intramuscular (i.m.) injection of plasmid DNA expression vectors. We previously demonstrated that this technique is an effective means of immunization to elicit both antibodies capable of conferring homologous protection and cell‐mediated immunity leading to cross‐strain protection against influenza virus challenge in mice. These results suggested that expression of viral proteins by muscle cells can result in the generation of cellular immune responses, including cytotoxic T lymphocytes (CTL). However, because DNA has the potential to be internalized and expressed by other cell types, we sought to determine whether or not induction of CTL required synthesis of antigen in non‐muscle cells and, if not, whether transfer of antigen to antigen‐presenting cells from muscle cells may be involved. In the present study, we demonstrate that transplantation of nucleoprotein (NP)‐transfected myoblasts into syngeneic mice led to the generation of NP‐specific antibodies and CTL, and cross‐strain protective immunity against a lethal challenge with influenza virus. Furthermore, transplantation of NP‐expressing myoblasts (H‐2^k) intraperitoneally into F1 hybrid mice (H‐2^d × H‐2^k) elicited NP CTL restricted by the MHC haplotype of both parental strains. These results indicate that NP expression by muscle cells after transplantation was sufficient to generate protective cell‐mediated immunity, and that induction of the CTL response was mediated, at least in part, by transfer of antigen from the transplanted muscle cells to a host cell.