CD8 + T cells are recognized as key players in exacerbation of ischemic stroke; however, the underlying mechanism in modulating the function of CD8 + T cells has not been completely elucidated. Here, we uncovered that FasL enhanced the cytotoxicity of CD8 + T cells to neurons after ischemic stroke. Inactivation of FasL specific on CD8 + T cells protected against brain damage and neuron loss. Proteomic analysis identified that PDPK1 functioned downstream of FasL signaling and inhibition of PDPK1 effectively reduced cytotoxicity of CD8 + T cells and improved ischemic neurological deficits. Taken together, these results highlight an intrinsic FasL-PDPK1 pathway regulating the cytotoxicity of CD8 + T cells in ischemic stroke.