Depletion of antigen-specific T cells directed toward MiHAg H4 and H60. Splenocytes from B6 mice were depleted with K^b-H4 tetramers plus K^b-H60 tetramers, or were mock depleted. Splenocytes (25 × 10⁶) were subsequently transferred into TCRα-deficient mice; 4 days later, mice received both H4 and H6 peptide vaccination. After vaccination (10 days), splenocytes were restimulated in vitro for 14 days in the presence of 5 × 10⁻⁴ μg/mL H4 (left panels), or 5 × 10⁻² μg/mL H60 peptide (right panels). Total alloreactive T-cell responses were assessed by intracellular IFNγ staining upon a 5-hour incubation with IFNγ-pretreated, CFSE-labeled BALB.B splenocytes (left plots), or with B6 control splenocytes (right plots). (A) Dot plots show the results of representative mice within each group with the percentage of IFNγ-producing CD8⁺ T cells depicted in the top right corner. (B) Graph shows the mean percentage of IFNγ-producing CD8⁺ T cells of 3 mice (H4: P = .015; H60: P = .084 ± SD).