Depletion of Treg by the Diphtheria Toxin System

Specific cell ablation by the diphtheria toxin (DT) system is widely used to analyze the in vivo function of target cells in mice. In this chapter, we describe the methods of depleting regulatory T cells (Tregs) systemically or selectively in the skin. Since it has been difficult to conclude the importance of tissue-residing Tregs with systemic Treg ablation, we sought to selectively deplete cutaneous Tregs to investigate their function in the skin without the depletion of Tregs in non-target organs. Here, we describe protocols for the depletion of Tregs by the DT system, and subsequent analysis of Tregs in the skin and skin-draining lymph node (dLN) by flow cytometry. This procedure of selective depletion of cutaneous Tregs can be applicable to other tissues and cells, to allow investigation of the role of tissue-resident cells in mice.