Figure 4. B7-H4 on AHSC inhibits cytokine producing CD8+T cells.
(A) Mock or B7-H4 siRNA transfected HSC were pulsed with gp33 peptide and cocultured with P14 CD8+ T cells for three days. The CD8+ T cells were then restimulated with plate bound anti-CD3/CD28 antibodies (1ug/ml) for 5 hours and intracellular IFNγ levels were analyzed. Thin black gate and grey numbers represent the percentage frequency of total IFNγ producing CD8+ T cells. Thick black gate and black numbers represent the frequency of high IFNγ producing CD8+ T cells. Plots are gated on CD3+CD8+ cells. Representative data is shown from two independent experiments (B) Fold difference in the frequency of total (left panel) or high MFI (right panel) IFNγ producing CD8+ T cells between B7-H4 knockdown and control HSC. Average and standard deviation is shown from three independent experiments. (C) CD8 + T cells cultured with anti-CD3/CD28 in the presence of B7-H4-Ig or control-Ig for three days were restimulated with plate coated anti-CD3/CD28 antibodies (1ug/ml) for 5 hours and intracellular IFNγ was measured. Representative data is shown from three independent experiments.