In recent years, the understanding of regulatory T cell (T_reg cell) biology has expanded considerably. Key observations have challenged the traditional definition of T_reg cells and have provided insight into the underlying mechanisms responsible for the development of autoimmune diseases, with new therapeutic strategies that improve disease outcome. This Review summarizes the newer concepts of T_reg cell instability, T_reg cell plasticity and tissue-specific T_reg cells, and their relationship to autoimmunity. Those three main concepts have changed the understanding of T_reg cell biology: how they interact with other immune and non-immune cells; their functions in specific tissues; and the implications of this for the pathogenesis of autoimmune diseases.