Foxp3+ regulatory T cells (Tregs) play essential roles in immune tolerance. In addition, Tregs have been shown to possess specialized functions in non-immune tissues such as visceral adipose tissue, skeletal muscle, and colonic lamina propria where Tregs contribute to tissue homeostasis. In this study, we discovered that Tregs accumulate in the brain one to two weeks after ischemic brain injury induced by a transient middle cerebral artery occlusion (MCAO). Analysis of gene expression profile revealed that the Tregs isolated from injured brain were different form other tissue Tregs. Tregs accumulated in the brain after ischemic injury expressed high levels of ST2 (IL-33 receptor) and Amphiregulin (Areg). Depletion of Tregs in DEREG mice resulted in the increase of the number of microglia as well as GFAP-positive reactive astrocytes (astrogliosis), and delayed neurological recovery. Rag-deficient mice also showed enhanced astrogliosis, and adoptive transfer of Tregs reduced astrogliosis. These data suggest that Tregs are neuroprotective through regulation of astrocytes. Areg may be involved in the regulation of astrogliosis, since administration of Areg reduced the number of activated astrocytes. FTY720 treatment reduced the number of Tregs in the brain, suggesting that Tregs were generated in the periphery and migrated into the brain. Brain Tregs expressed CCR6 and CCR8, and intraventricular administration of their ligands, CCL1 and CCL20 increased the infiltration of Tregs in the brain, and improved a neurological function. These results suggest that the accumulation of Tregs in the chronic phase after stroke is important for the neurological recovery and induction of brain-specific Tregs could be a useful therapy for alleviation of neurological symptoms.