T-cells are a part of the adaptive immune system. The function of T cells is dependent on T cell receptor-based activation, which is accompanied by cell growth, proliferation, and cytokine production. Jurkat cells are an immortalized human cell line derived from the T cell of a patient with acute leukemia. This cell line has been used for understanding many T cell receptor signaling and activation processes. Here we used this cell line to decipher the role of glutamine as a nutrient during the activation process and its cross-talk with the Notch1 signaling pathway. The activation of Jurkat T cells by phytohaemagglutinin (PHA)/phorbol myristate acetate (PMA) leads to a suppression of canonical Notch signaling pathway along with an increase in IL2 production in the presence of glutamine. This also increased the glutamine consumption by cells. Upon removal of glutamine, IL2 production decreased as was observed for murine-derived T effector cells. Interestingly, we found that constitutive expression of Hey1, the downstream effector of Notch pathway, prevented the surge in IL2 secretion upon activation, but the IL2 secretion steadily increased in glutamine-free condition. This observation shows that Notch signaling pathway effectors can modulate the cytokine secretion depending on the presence or absence of exogenous glutamine. This effect may be exploited in studying the modulation of T-cell activation in tumor microenvironment where there is a competition for nutrients between the proliferating tumor cells and the activated T cells.