CKLiK297-321 abrogates fMLP-induced migration and inhibits fMLP-induced β2-integrin activation. (A) Migration of granulocytes was monitored by time-lapse analysis. Cells were first allowed to attach on albumin-coated coverslips for 15 minutes. Granulocytes were incubated as indicated with 50 μM CKLiK297-321 or 50 μM scrambled tat control peptide. Neutrophil migration was induced by fMLP (10-8 M) and imaged every 20 seconds for 10 minutes. Migration tracks of individual cells are shown per cell (top row) and centered (bottom row). (B) Average migration speed of at least 3 independent experiments are calculated and expressed as micrometers per minute ± SEM. (C) Granulocytes were preincubated with CKLiK297-321 or with scrambled tat control peptide in the presence of control antibody (W6/32) or blocking antibody against β2-integrins (IB4) as indicated. Preincubated granulocytes were added to wells containing buffer or fMLP (10-8 M) and ICAM-1-coated beads. After 15 minutes at 37°C the cells were washed, and the percentage of cells that were positive for beads was determined by flow cytometry. Depicted is the average of 3 experiments with SEM.
