There is emerging evidence that IFNβ, which has been discovered as a major effector cytokine of the immune response against viral infections, can exert multiple functions in host defense and homeostasis. T cells are the major component of adaptive immunity and respond to IFNβ released by innate immune cells, resulted in regulating host immune response against viral infection and cancer. However, the molecular mechanisms by which IFNβ regulates T cell response in human are largely elusive. Answering this question is critical to better understand T cell response to IFNβ and to identify therapeutic target for individual patients who are suffering with infectious diseases, autoimmunity, and cancer. Here, we performed unbiased high-resolution transcriptional profiling of human T cells and constructed an IFNβ-inducible transcriptional regulatory network in human T cells. Our unbiased approach to investigate the kinetics of global gene expression allow us to identify novel IFNβ-responsible genes that have been overlooked by single snapshot-based exploration. We found some of the genes are important to induce expression of exhaustion markers, suggesting that our method could dissect the IFNβ-induced exhaustion program. This high-resolution gene expression profiling will provide a platform that enables us to connect each “dots”, which are already known as individual IFNβ responses, providing the systematic understanding of the molecular network of immune reaction to viral infection, autoimmune and cancer.