GVHD occurs independent of accumulation of H60-specific T cells within the target organs at the time of disease. Splenocytes from female B6 mice were either depleted with K^b-H4-and K^b-H60 tetramers or mock depleted. Subsequently, either 25 × 10⁶ H4/H60-depleted T cells (top row), 25 × 10⁶ mock-depleted T cells (middle row), or 2 × 10⁶ mock-depleted splenocytes (bottom row) plus 5 × 10⁶ T-cell–depleted BM cells were transferred into lethally irradiated male BALB.B recipients (n = 6 in all groups). After onset of clinical GVHD (•) or at day 21 (○) blood, spleens, and livers were harvested to determine H60-specific or OVA-specific T-cell responses by intracellular IFNγ staining. Each circle represents 1 mouse; bars indicate averages.