Fig. 1

The generation of recombinant proteins and binding conformations of CD80/CD86-Ig and functional analysis of cytotoxic T-lymphocyte antigen 4 (CTLA-4)-Ig. CTLA-4-Ig, CD80-Ig, and CD86-Ig were produced in Expi293F cells that were transfected with expression vectors. a The purification of CTLA-4-Ig, CD80-Ig, and CD86-Ig was confirmed using SDS-PAGE. b The binding of CTLA-4-Ig against CD80-enhanced green fluorescent protein (EGFP) or CD86-EGFP expressing Cos-7 cells was confirmed using flow cytometry. c The binding of rabbit IgG, CD80-Ig, and CD86-Ig against CTLA-4-EGFP expressing CHO cells was confirmed using flow cytometry. d Peripheral blood mononuclear cells (PBMCs) (n = 11) were isolated from healthy cattle and cultured with 10 nM rabbit IgG or CTLA-4-Ig in the presence of Staphylococcus enterotoxin B (SEB). The cell culture supernatant was harvested after 7 days, and an ELISA was used to measure the inhibited interferon-gamma (IFN-γ) concentration. The bars indicate the average of each group. The statistical comparisons between each group were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at p < 0.01