FIG. 4.
Incubation of bovine PMNs with inflammatory cytokines enhances LKT binding. Freshly isolated bovine PMNs (1 × 106 cells/ml) were incubated with recombinant bovine IL-1β (50 ng), recombinant human TNF-α (50 ng), recombinant bovine IFN-γ (50 ng), or medium (control) for 15 min (A) or 60 min (B) at 37°C. The cells were then incubated for 10 min in an ice bath with biotinylated LKT (1 U) and washed. Extra-avidin-FITC was added and incubated for 20 min on ice. The stained cells were washed, fixed with paraformaldehyde, and analyzed by flow cytometry (10,000 cells were scored for green fluorescence). The data are the means ± standard errors of the means for four independent experiments. An asterisk indicates that the data for stimulated cells was statistically significantly different from the data for control cells (P < 0.05).