Regulatory T (Treg) cells have emerged as a central control point in the modulation of various immune responses, including autoimmune responses and immunity to transplants, allergens, tumors, and infectious microbes. The immune system has evolved complex processes to ensure tolerance to autoantigens while preserving the potential to mount and maintain life-long humoral and cellular immune responses against invading pathogens. In this review, we summarize research showing that naturally occurring (nTreg) and induced Treg (iTreg) cell subsets, and in particular CD4+Foxp3+ Treg cells, are critical in the control of immune responses in rodents and humans. We also discuss the cellular and molecular factors that affect CD4+Foxp3+ Treg cell development, homeostasis, and function and consequential immunity to self and non-self antigens. Recent studies have shed light in our understanding of the development of novel methods of autoimmune disease prevention and treatment via enhancing and re-establishing Treg-mediated dominant control over self-reactive T cells in animal models and humans.