Objective: To study NF-κB activation during the early phase of T cell reaction due to CD3/CD28 co-stimulation and the effect of arsenic trioxide on NF-κB. Methods: Hut-78 cells were stimulated with CD3/CD28 monoclonal antibodies(10 μg/ml and 5 μg/ml, respectively) in 30 min. TransAM was employed to investigate p65 expression in nuclear extracts and the effect of arsenic troxide. Results: The level of p65 in nuclear extracts of Hut-78 cells was much lower during the CD3/CD28 co-stimulation in 30 min, however arsenic trioxide seemed to inhibit the effect of CD3/CD28 co-stimulation. And the trend was nearly the same in whole cell lysates. Conclusion: NF-κB is suppressed during the early phase of CD3/CD28 co-stimulation in human T cells.