Functional and ultrastructural analysis of annexin A1 and its receptor in extravasating neutrophils during acute inflammation.
The purpose of this study was twofold: to reveal cellular events associated with the protective role of endogenous annexin A1 (AnxA1) in inflammation and to highlight the potential involvement of members of the formyl peptide receptor (Fpr) family in this process. We found that wild-type, AnxA1-null, and Fpr1-null mice all displayed an intense neutrophil recruitment into the peritoneal cavity as assessed 4 hours after carrageenin injection, and that this recruitment was most pronounced in AnxA1-null mice. In addition, this cell influx could be inhibited by the AnxA1 pharmacophore peptide, Ac2-26, in wild-type, AnxA1-null, and Fpr1-null mice, but was restored when co-treated with the pan-receptor antagonist Boc2. Using the LacZ gene reporter assay, an enhancement of AnxA1 gene promoter activity in extravasated neutrophils was evident in AnxA1-null mice; again this response was reduced after peptide treatment. The lack of functional involvement of Fpr1 prompted us to monitor the structurally related receptor Fpr2. We report, for the first time, the ultrastructural immunocytochemical co-localization of Fpr2 with AnxA1 in neutrophils that migrate into the mesenteric microcirculation and extravasate into the peritoneal fluid. Collectively, these data provide in vivo support to the hypothesis that endogenous AnxA1 is an essential effector of endogenous anti-inflammation and provide an ultrastructural indication that this mediator interacts with Fpr2 in murine neutrophils. We believe that these findings could significantly affect the development of novel therapeutics, which are modeled after the anti-migratory actions of AnxA1.
Gastardelo
Thaís Santana
TS
Post-Graduation in Morphology, Federal University of São Paulo, Paulista School of Medicine (EPM), São Paulo, Brazil.
Damazo
Amílcar Sabino
AS
Dalli
Jesmond
J
Flower
Roderick J
RJ
Perretti
Mauro
M
Oliani
Sonia Maria
SM
D000818:Animals; D017305:Annexin A1 / Q000378:metabolism* / Q000648:ultrastructure; D002634:Chemotaxis, Leukocyte / Q000276:immunology*; D015870:Gene Expression; D007150:Immunohistochemistry; D007249:Inflammation / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D044042:Receptors, Formyl Peptide / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017305:Annexin A1; C469988:Fpr1 protein, mouse; D044042:Receptors, Formyl Peptide; C481578:formyl peptide receptor 2, mouse