Annexin 1 and its bioactive peptide inhibit neutrophil-endothelium interactions under flow: indication of distinct receptor involvement

RPG Hayhoe, AM Kamal, E Solito, RJ Flower… - Blood, 2006 - ashpublications.org
RPG Hayhoe, AM Kamal, E Solito, RJ Flower, D Cooper, M Perretti
Blood, 2006ashpublications.org
We have tested the effects of annexin 1 (ANXA1) and its N-terminal peptide Ac2-26 on
polymorphonuclear leukocyte (PMN) recruitment under flow. Differential effects of the full-
length protein and its peptide were observed; ANXA1 inhibited firm adhesion of human
PMNs, while Ac2-26 significantly attenuated capture and rolling without effect on firm
adhesion. Analysis of the effects of ANXA1 and Ac2-26 on PMN adhesion molecule
expression supported the flow chamber results, with Ac2-26 but not ANXA1 causing l …
We have tested the effects of annexin 1 (ANXA1) and its N-terminal peptide Ac2-26 on polymorphonuclear leukocyte (PMN) recruitment under flow. Differential effects of the full-length protein and its peptide were observed; ANXA1 inhibited firm adhesion of human PMNs, while Ac2-26 significantly attenuated capture and rolling without effect on firm adhesion. Analysis of the effects of ANXA1 and Ac2-26 on PMN adhesion molecule expression supported the flow chamber results, with Ac2-26 but not ANXA1 causing l-selectin and PSGL-1 shedding. ANXA1 and its peptide act via the FPR family of receptors. This was corroborated using HEK-293 cells transfected with FPR or FPRL-1/ALX (the 2 members of this family expressed by human PMNs). While Ac2-26 bound both FPR and FPRL-1/ALX, ANXA1 bound FPRL-1/ALX only. ANXA1 and Ac2-26 acted as genuine agonists; Ac2-26 binding led to ERK activation in both FPR- and FPRL-1/ALX-transfected cells, while ANXA1 caused ERK activation only in cells transfected with FPRL-1/ALX. Finally, blockade of FPRL-1/ALX with a neutralizing monoclonal antibody was found to abrogate the effects of ANXA1 in the flow chamber but was without effect on Ac2-26-mediated inhibition of rolling. These findings demonstrate for the first time distinct mechanisms of action for ANXA1 and its N-terminal peptide Ac2-26.
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