American Society of Hematology

Figure 2.

$Figure 2. In vitro analysis of donor splenocyte depletion. (A) Following tetramer depletion, pre- and postdepletion fractions were stained with LYL8 and HSV8 tetramers. The percent retention of tetramer-positive cells after depletion was determined as follows: 100 × (% postdepletion / % predepletion). These data are from 4 independent experiments and represent the mean plus or minus SEM. (B) B6 splenocytes from pre- and postdepletion fractions were incubated with anti-CD3, CD20, CD11b, GR-1, and NK1.1 fluorescently labeled antibodies. Percentage is based on positive gating on single parameter histograms. (C) B6 splenocytes from pre- and postdepletion fractions were coincubated with FITC-labeled antibodies against the members of the Vβ family represented on the x-axis and PE-anti-CD3 antibodies. Percentage is based on positive gating on single parameter histograms of CD3+ cells. These data are representative of 2 repeated experiments. (D) B6 splenocytes from pre- and postdepletion fractions were incubated in MLRs with either mHAg (BALB.B) or MHC (BALB/c) mismatched irradiated splenocytes. Each bar is representative of the mean of quadruplicates plus or minus SEM.$

In vitro analysis of donor splenocyte depletion. (A) Following tetramer depletion, pre- and postdepletion fractions were stained with LYL8 and HSV8 tetramers. The percent retention of tetramer-positive cells after depletion was determined as follows: 100 × (% postdepletion / % predepletion). These data are from 4 independent experiments and represent the mean plus or minus SEM. (B) B6 splenocytes from pre- and postdepletion fractions were incubated with anti-CD3, CD20, CD11b, GR-1, and NK1.1 fluorescently labeled antibodies. Percentage is based on positive gating on single parameter histograms. (C) B6 splenocytes from pre- and postdepletion fractions were coincubated with FITC-labeled antibodies against the members of the Vβ family represented on the x-axis and PE-anti-CD3 antibodies. Percentage is based on positive gating on single parameter histograms of CD3⁺ cells. These data are representative of 2 repeated experiments. (D) B6 splenocytes from pre- and postdepletion fractions were incubated in MLRs with either mHAg (BALB.B) or MHC (BALB/c) mismatched irradiated splenocytes. Each bar is representative of the mean of quadruplicates plus or minus SEM.

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