Abstract Autoimmune diseases occur when there is misregulation within the immune system. Regulatory T cells (Tregs) are critical for the regulation of this immune response, however, little is known about their own regulation. The induction and activation of Tregs have been shown to alleviate symptoms of autoimmune disease such as human multiple sclerosis (MS). Our lab has identified Tregs that are activated by the random amino acid copolymer YFAK. These YFAK-specific Tregs can reduce the symptoms of experimental autoimmune encephalomyelitis (EAE), the animal model of MS, more effectively than YEAK (Copaxone), a drug currently used in MS therapy. We have generated both YFAK-specific T cell lines as well as retrogenic mice expressing YFAK-specific T cell receptors (TCR) in order to study the regulation and activation of these beneficial Tregs. We find that these YFAK-specific Tregs will proliferate upon stimulation with glucococorticoid-induced TNF receptor ligand (GITRL). In addition, these Tregs can be activated to produce high levels of anti-inflammatory cytokines, and are immunosuppressive against conventional T cells. We plan to further characterize these YFAK-specific Tregs, as well as to utilize the TCR retrogenic mice in in vivo functional experiments. These studies will help to further explore the use of YFAK-specific Tregs in the prevention of autoimmune diseases such as multiple sclerosis.