Prevention of in vitro neutrophil adhesion to endothelial cells through shedding of L-selectin by C-reactive protein and peptides derived from C-reactive protein

C-Reactive protein (CRP), the classic acute-phase reactant in humans, diminishes accumulation of neutrophils at inflammatory sites, To evaluate the underlying mechanisms, we have studied whether CRP and peptides derived from CRP could affect the first step of neutrophil extravasation, the L-selectin-mediated interaction of neutrophils with endothelial cells, CRP markedly attenuated attachment of human neutrophils to cultured LPS-activated human coronary artery and pulmonary microvascular endothelial cells with apparent IC50 values of 20 and 22 mu g/ml, respectively, At similar concentrations, CRP rapidly downregulated the expression of L-selectin on the neutrophil surface, whereas it failed to affect expression of CD11b and CD45 or to induce granule enzyme release, The loss of L-selectin was due to cleavage and shedding of the molecule from the cell surface, as quantitated by the soluble form of L-selectin in cell-free supernatants, The effects of CRP could be prevented by an anti-CRP antiserum and by KD-IX-73-4, which inhibits shedding of L-selectin. Inhibition of adhesion with CRP was additive with function-blocking anti-E-selectin and anti-CD18 antibodies, but was not additive with anti-L-selectin antibody, Neutrophil attachment and L-selectin expression were also diminished by CRP peptides 174-185 and 201-206, but not peptide 77-82, albeit these peptides showed a weaker inhibitory effect than the parent protein, These studies indicate a specific activation-independent action of CRP and CRP peptides 174-185 and 201-206 on expression of L-selectin, and suggest that by attenuating neutrophil adhesion to the endothelium and consequently neutrophil traffic into tissues, native CRP and peptides 174-185 and 201-206 may be major mechanisms to attenuate or limit the inflammatory response.