Figure 7. HLA class II tetramers detect S-Ag-responsive cells in PBMC of uveitis patient.

PBMC from HLA-DR3 or from HLA-DR4 uveitis patients were stimulated with peptide N282 for 7 days as described in Methods. Cells were re-stimulated on day 7 and expanded in IL-2 containing medium. On day17 of culture cells were stained with HLA-DR3 tetramer (PE) loaded with N282 peptide (NRERRGIALDGKIKHE) or with unloaded HLA-DR3 tetramer (PE) followed by anti-CD4 (APC) and anti-CD3 (FITC). Dead cells were excluded from analysis (7AAD). Percentages of tetramer⁺ CD3⁺ cells are shown after gating on CD4⁺ 7-AAD⁻ cells.