The initiation step of cell-mediated immune responses in the central nervous system (CNS) involves the trafficking of the antigen-specific T cells into the brain. To study this trafficking, we developed an in vivo system for studying antigen-specific responses in the CNS. In this assay, T cell receptor (TCR) transgenic mice having 95% of T cells specific for a defined antigen-pigeon cytochrome c (PCC) were cannulated intraventricularly for PCC antigen infusion and cerebrospinal fluid (CSF) sampling. Upon PCC infusion into the CNS, the number of alpha/beta TCR(+) Vbeta3(+) Mac1(-) cells in the CSF was characterized. We found that infusion of antigen into the CSF induced an increased number of antigen-specific T cells in the CNS and activation of antigen-specific T cells in the peripheral blood. Hence, the drainage of CNS antigen into the periphery might play an important role in sustaining autoimmune reactivity in CNS inflammatory diseases.