Figure 3
Characterization of Boch1D10F1. A Reactivity of Boch1D10F1 with BoPD-1-myc cells. BoPD-1-myc cells were stained with Boch1D10F1 in serial dilutions (10 μg/mL to 100 pg/mL) and analyzed by flow cytometry. B Blockade of PD-1/PD-L1 binding by Boch1D10F1. BoPD-1-Ig was preincubated with 5D2 and Boch5D2 and incubated with BoPD-L1-EGFP cells. BoPD-1-Ig bindings were evaluated by flow cytometry. Each curve represents the relative binding of BoPD-1-Ig preincubated with Boch1D10F1, Boch5D2, 1D10F1, and 5D2 compared with no-antibody control. Bovine IgG (for Boch1D10F1 and Boch5D2), rabbit IgG (1H4L), and rat IgG2a controls (for 5D2) were used as negative controls. C, D Effect of PD-1/PD-L1 blockade on the IFN-γ response. PBMCs of BLV-infected cattle (n = 13) were cultured with Boch1D10F1 and Boch5D2 (10 μg/mL) or bovine IgG control in the presence of FLK-BLV antigen (2%) for 6 days. IFN-γ production for each animal was measured by ELISA in duplicate. Bars indicate the group median response. Significant differences between each group were determined using Wilcoxon signed-rank test Friedman test. **p < 0.01.