Introduction: The herpes simplex viruses (HSV) cause a variety of clinical illnesses that are painful and often distressing. To control the infection, the development of an effective vaccine that prevents or reduces the primary and recurrent infections would be of great significance. Chronic and acute exercise training were used as a vaccine adjuvant in some studies. The purpose of this study was to assess adjuvant effects of short term endurance exercise training on regulatory T cells in HSV-1 vaccine model. Methods & Materials: 24 BALB/C mice were divide into 3 groups: control, vaccine and vaccine plus short term endurance exercise training. TGF-β levels as index of reglulatory T cells activation were determined in spleen cell culture with ELISA method. One-way ANOVA were used for statistical analysis. Results: The result of this study indicated that there are significant differences between group in reglulatory T cells activation (P<0.05). Tukey post-hoc test results showed significant differences between control and vaccine groups with vaccine plus short term endurance exercise training group (P<0.05). Conclusion: Results of current study showed that reglulatory T cells activation increased following using short term endurance exercise training as an adjuvant. These results suggested that coupling short term moderate exercise training with vaccination may enhance vaccine efficacy. Further studies are in order to determine effects of different kind of exercise as an adjuvant in different types of vaccine can be considerate.