We have found previously that early activated T cells show higher activities and antitumor effects than late activated T cells upon their adoptive transfer. However, the exact mechanism is not clear. This study aimed to investigate the impact of polyclonal activation on the metabolic signature of T cells from naïve mice. Splenocytes of Swiss albino mice were harvested and cultured at 2.5 × 10⁶/ml in  T25 flasks in presence of (Con-A) and IL2 for 24h or 72h Then, cells were harvested and processed to determine  activities of hexokinase (HK), phosphofructokinase (PFK), lactate dehydrogenase (LDH) and glucose 6 phosphate dehydrogenase(G6PD).  The obtained data showed that activity of enzymes in activated cellswas increased after 24h and 72h activation as compared to naïve cells, when  and supernatant their level at 72h activation was higher than 24h activation. Comparison of enzymes activity between cells and supernatant showed higher activity of HK, G6PD and LDH in cells, While PFK activity was higher  in supernatant after 72h.these results indicate that Upon the early activation, naïve T cells have higher glycolytic pathway than late activation. These results have a great implication in adoptive T cell therapy