Previous studies have found the close correlation between epitope density and epitope-specific response, which have shown that high epitope density in a single recombinant protein molecule significantly enhances the humoral response and protective immunity. However, it has not been determined whether this kind of high epitope density could also significantly influence T cell response. Based on this, a series of recombinant DNA and proteins were designed and prepared. Each molecule consists of various copy numbers of the V3 CTL epitope on HIV-1 gp120 (one, two, four and eight copies). Our results show clearly that different V3-epitope densities in just one single DNA or protein molecules have respectively different effects on the number and activity of both primary and memory T cells. Interestingly, this effect is more complex than that on the B cells: epitope density in one plasmid or protein antigen affects the number, not the cytotoxic avidity, of primary CD8+ T cells, but affects both the number and cytotoxic avidity of memory CD8+ T cells. It indicates epitope density in the antigen is an important consideration to optimize T cell response induction and may facilitate the development of effective T cell-based anti-virus vaccines.