The objective of this study is to generate dendritic cell (DC) vaccines by exposing DCs to C6 glioma cancer cell antigenic (tumor) peptides following the exposure of C6 cells to photodynamic therapy (PDT) and acid elution. Effects of these DCs on host immunity were assessed by measuring cytokine induction (following adaptive transfer into rats) and assessing DC-induced cytotoxic T lymphocyte (CTL)-mediated lysis of C6 target cells. Precursor dendritic cells were purified from rat bone marrow and matured in vitro. C6 cells were stimulated with PDT, and adherent cells were acid-eluted to obtain cell surface antigens, whole cell antigens were also isolated from supernatants. C6 cells not stimulated with PDT were also used to isolate antigens by acid elution or freeze-thaw methods for comparison purposes. The isolated antigens from the respective purification methods were used to sensitize DCs for the generation of DC vaccines subsequently transferred into SD rats. Following adoptive transfer, the changes in interleukin (IL)-12, IL-10, and TNF-α expression were measured in rat serum by ELISA. CTL-mediated lysis was assessed using the MTT assay. PDT-generated antigens further purified by acid elution had the greatest stimulatory effect on DCs based on the elevated serum IL-12 and TNF-α levels and decreased serum IL-10 levels. CTL activity in this group was also highest (percent lysis 95.5% ± 0.016) compared with that elicited by PDT-supernatants, acid elution, and freeze-thawing (or the control group), which had 90.2% ± 0.024, 73.3% ± 0.027, 63.6% ± 0.049, or 0.4% ± 0.063 lysis, respectively. PDT significantly enhanced tumor cell immunogenicity. These data suggested that DC vaccines prepared by treating tumor cells with PDT to generate antigen-specific CTL responses can be developed as novel cancer immunotherapeutic strategies.