Figure 6.

Severe EAU induced with a T cell line that is stably polarized to the Th1 phenotype. (A and B) After 48 h of stimulation with p161–180, the indicated number of cells was infused into naive B10RIII mice. Shown are average scores (A) and representative histopathology. (B) Inset shows the typical inflammatory infiltrate. Bar, 0.25 mm (0.02 mm in inset). (C) ELISA analysis of cytokines in supernatants of the stimulated cells. (D and E) Intracellular cytokine staining after stimulation with Ag for 24 h in neutral conditions (D) or for 5 d under Th17-inducing conditions (Ag plus IL-23 or Ag plus IL-23 plus anti–IFN-γ; E). (F) “Parking” of Th1 cells in allotype-congenic recipient mice. After stimulation with p161–180 in the presence of irradiated APCs, the T cell line was infused into naive Thy1.1 x Thy1.2 heterozygous recipients (2 × 10⁶/mouse). Note heterozygous Thy1.1/2⁺ recipient cells versus Thy1.2⁺ donor cells by FACS analysis. Thy1.2 single-positive cells were sorted out after 90 h, stimulated with IRBP peptide 161–180 for 24 h (with PMA-ionomycin and brefeldin A added during the last 4 h), and stained for intracellular IL-17 and IFN-γ. Representative experiment of two with five mice per group.