Effects of inhibition of β2-integrin function on adhesion to TNF-treated EC and migration into subendothelial matrix.

<p>HUVEC were stimulated with 100U/ml TNF for 4 h and neutrophils were added and allowed to adhere for 10 min before non-adherent cells were washed off and data collected at 15 min, 1 h, 3 h and 24 h. A,D: Total adherent neutrophils after 15min (% of those added). B,E: Neutrophil transmigration after 1 hour (% of those adherent). C,F: Average depth penetrated by the neutrophils that had transmigrated. In A-C neutrophils were untreated, pre-treated with antibody against β2-integrin, or antibody was added after 15min. InD-F, neutrophils were untreated, or gels were loaded with a small molecule inhibitor of β2-integrin (CT7010). G: Fluorescence intensity distribution for neutrophils retrieved from the gel at 3h when antibody against β2-integrin was added after 15min, assessed by flow cytometry. In A and B, ANOVA showed a significant effect of treatment (p<0.01 in each case); * = p<0.05, ** = p<0.01 by Dunnett test compared to untreated. In D, ** = p<0.01 by paired t-test compared to untreated. In C and F, ANOVA showed significant effect of time (p<0.01 in each case) but not treatment. Data are mean ± SEM from 4 experiments (A-C) or 5 experiments (D-F).