Regulatory T cells inhibit cellular immunity and represent an obstacle for the development of cancer immunotherapy. The understanding of Treg cellular biology has exponentially increased during the last 10 years, driven primarily by elegant in vivo studies of mouse models systems and in vitro studies of human cells. Numerous clinical strategies are under active investigation to achieve Treg depletion or inhibition in patients with cancer, including low-dose cyclophosphamide and interleukin-2 or anti–interleukin-2R immunotoxins. To date, only modest results have been reported in patients. Our preliminary data suggest that the antihuman CD25 monoclonal daclizumab may be useful as an alternative approach for in vivo Treg depletion, but the mechanism of action of this effect remains to be elucidated. Certain immune modulatory agents may indirectly affect Tregs in patients with cancer but not necessarily in the desired direction for the therapeutic setting. More sophisticated techniques that have become available for Treg analysis in patients will assist in this important translational effort.