Effects of inhibition of integrin function on entry of neutrophils into collagen gels and depth of penetration induced by fMLP.

<p>Collagen gels were formed and loaded with (A) or confocal microscopy (B). B. Representative images of neutrophil nuclei (i) or intracellular bioparticles at 60min (ii) analysed by confocal microscopy., before neutrophils were added and allowed to settle and migrate for 30 and 60 min. A,C,E: Proportion of added neutrophils entering the gel. B,D,F: Average depth penetrated by the neutrophils that had entered. In A, B, neutrophils were untreated, or treated with antibody against β2-integrin or a small molecule inhibitor (CT7010). In C, D, neutrophils were untreated, or treated with antibody against β1 or against β3-integrin, or with RGDS peptide. In E, F, neutrophils were untreated, or pre-treated with antibodies against β1-, β2- and β3-integrin, or with a combination of non-specific IgG with matching isotypes. In A and E, ANOVA showed a significant effect of treatment on entry into the gel (p< 0.01 in each case); in addition, Dunnett test for combined time points showed significant effects of antibodies against integrin or CT7010 (* = p<0.05; ** = p<0.01). In F, ANOVA showed a significant effect of treatment on depth of penetration (p<0.01); in addition, Dunnett test for combined time points showed a significant effect of the combination of antibodies against integrins (* = p<0.01). Data are mean ± SEM from 4 experiments, except for CT7010 tested on 3 occasions.