For intracellular cytokine stimulation assays, lymphocytes were isolated from brain and spleen, cultured in DMEM/10% FBS for 5 h at 37˚C with or without 1 μM LT359 peptide [84] , stained with Fixable Viability Dye, anti-CD8α, and anti-CD44, then permeabilized and fixed in FoxP3 buffer fixation and permeabilization solutions.