Leukocyte adhesion in the flow chamber. Leukocyte adhesion (mean ± SEM of cells/mm2) under shear (1 dyne/cm2) was observed in the microflow chamber assay using myelomonocytic cells isolated from LysEGFP mice (A) or human PMNs (B) at a final concentration of 0.25 × 106 cells. rmP-selectin (2 μg/mL), rmCXCL1 (KC, 5 μg/mL), and sRAGE (1 μg/mL) were immobilized in flow chambers for experiments with murine neutrophils (A). rhP-selectin (4 μg/mL), rhCXCL8 (IL-8, 10 μg/mL), and sRAGE (4 μg/mL) were used in experiments with human neutrophils (B). Blockade of Mac-1 was achieved by preincubating the cell suspensions for 20 minutes with Mac-1–blocking mAbs Tib128 (antimouse) or ICRF44 (antihuman; 10 μg/106 cells), respectively. LFA-1 was blocked by mAb Tib217 (antimouse) or HI111 (antihuman; 10 μg/106 cells), respectively. Data are mean ± SEM from at least 4 flow chamber experiments per group. *Significant differences in adhesion to control PMNs perfused flow chambers coated with P-selectin, rmCXCL1 (A) or rhCXCL8 (B), and sRAGE (P < .05). n.a. indicates not assessed.
