Fig. 1.

The binding and blocking abilities of anti-bovine cytotoxic T lymphocyte antigen 4 (CTLA-4) mAb (4C2-D9). Bovine CTLA-4-EGFP-expressing cells was incubated with anti-bovine CTLA-4 mAbs (4C2-D9 and 4G2-A3) to assess binding and blocking ability. (A) The binding ability of anti-bovine CTLA-4 mAbs was measured using flow cytometry. (B and C) Concentration-dependent blocking effect of the binding of anti-bovine CTLA-4 mAbs (4C2-D9 and 4G2-A3) to CTLA-4/CD80 (B) and CTLA-4/CD86 (C). CTLA-4-EGFP-expressing cells were preincubated with 4C2-D9, 4G2-A3, or a mouse IgG₁ control (1.25, 2.5, 5, 10, and 20 μg/ml). Ig binding was detected using flow cytometry. Each point indicates the mean value of the relative MFI of three independent experiments (relative to the control without antibody; error bar, SEM). Tukey’s test was used for statistical analysis (*P<0.05) between groups treated with the same concentration of 4G2-A3 and mouse IgG₁. ‡P<0.05, between the groups treated with the same concentration of 4C2-D9 and mouse IgG₁. †P<0.05, between the groups with the same concentration of 4C2-D9 and 4G2-A3.