Activated T Cells Promote the Activation of Uveitogenic T Cells and Exacerbate EAU Development

METHODS. T cells were isolated from B6 mice immunized with the uveitogenic peptide IRBP1–20 and T cells from immunized TCR/ mice. Resting T cells were prepared by culture of separated T cells in cytokine-free medium for 3 to 5 days, when they showed downregulation of CD69 expression. Activated T cells were prepared by incubating resting T cells with antiTCR (GL3) for 2 days. Responder T cells were cocultured with immunizing antigen and antigen-presenting cells. The numbers of antigen-specific T cells expressing IL-17 or IFNwere determined by intracellular staining followed by FACS analysis after stimulation, with or without the addition of purified T cells. The cytokines in the culture medium were measured by ELISA.