Evidence for an anti-inflammatory loop centered on polymorphonuclear leukocyte formyl peptide receptor 2/lipoxin A4 receptor and operative in the inflamed microvasculature

• Published in: The Journal of immunology (1950) 2011-04, Vol.186 (8), p.4905-4914
• Main Authors: Brancaleone, Vincenzo, Dalli, Jesmond, Bena, Stefania, Flower, Roderick J, Cirino, Giuseppe, Perretti, Mauro
• Format: Article
• Language: English
• Subjects: Animals; Annexin A1 - genetics; Annexin A1 - immunology; Annexin A1 - metabolism; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Blotting, Western; Cell Membrane - metabolism; Dose-Response Relationship, Drug; Flow Cytometry; Humans; Inflammation - genetics; Inflammation - immunology; Inflammation - prevention & control; Inflammation Mediators - immunology; Inflammation Mediators - metabolism; Lipoxins - pharmacology; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Microvessels - immunology; Microvessels - metabolism; Microvessels - pathology; Neutrophils - drug effects; Neutrophils - immunology; Neutrophils - metabolism; Oligopeptides - pharmacology; Peptide Fragments - pharmacology; Protein Transport - drug effects; Receptors, Formyl Peptide - immunology; Receptors, Formyl Peptide - metabolism; Receptors, Lipoxin - immunology; Receptors, Lipoxin - metabolism; Time Factors
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.1003145

The importance of proresolving mediators in the overall context of the resolution of acute inflammation is well recognized, although little is known about whether these anti-inflammatory and proresolving molecules act in concert. In this article, we focused on lipoxin A(4) (LXA(4)) and annexin A1 (AnxA1) because these two very different mediators converge on a single receptor, formyl peptide receptor type 2 (FPR2/ALX). Addition of LXA(4) to human polymorphonuclear leukocytes (PMNs) provoked a concentration- and time-dependent mobilization of AnxA1 onto the plasma membrane, as determined by Western blotting and flow cytometry analyses. This property was shared by another FPR2/ALX agonist, antiflammin-2, and partly by fMLF or peptide Ac2-26 (an AnxA1 derivative that can activate all three members of the human FPR family). An FPR2/ALX antagonist blocked AnxA1 mobilization activated by LXA(4) and antiflammin-2. Analysis of PMN degranulation patterns and phospho-AnxA1 status suggested a model in which the two FPR2/ALX agonists mobilize the cytosolic (and not the granular) pool of AnxA1 through an intermediate phosphorylation step. Intravital microscopy investigations of the inflamed mesenteric microvasculature of wild-type and AnxA1(-/-) mice revealed that LXA(4) provoked leukocyte detachment from the postcapillary venule endothelium in the former (>50% within 10 min; p < 0.05), but not the latter genotype (∼15%; NS). Furthermore, recruitment of Gr1(+) cells into dorsal air-pouches, inflamed with IL-1β, was significantly attenuated by LXA(4) in wild-type, but not AnxA1(-/-), mice. Collectively, these data prompt us to propose the existence of an endogenous network in anti-inflammation centered on PMN AnxA1 and activated by selective FPR2/ALX agonists.