Human neutrophil migration in vitro induced by secretory phospholipases A(2): a role for cell surface glycosaminoglycans

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Gambero, A.
Landucci, ECT
Toyama, M. H.
Marangoni, S.
Giglio, JR
Nader, H. B.
Dietrich, C. P.
De Nucci, G.
Antunes, E.
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The purpose of this study was to examine the ability of type I- (porcine pancreas and Naja mocambique mocambique venom), type II- (bothropstoxin-I, bothropstoxin-II, and piratoxin-I). and type III- (Apis mellifera venom) secretory phospholipases A(2) (sPLA(2)s) to induce human neutrophil chemotaxis, and the role of the cell surface proteoglycans, leukotriene B-4 (LTB4), and platelet-activating factor (PAF). in mediating this migration. the neutrophil chemotaxis assays were performed by using a 48-well microchemotaxis chamber. Piratoxin-I, bothropstoxin-I. N. m. mocambique venom PLA(2) (10-1000 mug/mL each), bothropstoxin-II (30-1000 mug/mL), porcine pancreas PLA(2) (0.3-30 mug/mL), and A. mellifera venom PLA(2) (30-300 mug/mL) caused concentration-dependent neutrophil chemotaxis. Heparin (10-300 U/mL) concentration-dependently inhibited the neutrophil migration induced by piratoxin-I, bothropstoxin-II, and N. in. mocambique and A. mellifera venom PLA(2)s (100 mug/mL each), but failed to affect the migration induced by porcine pancreas PLA(2). Heparan sulfate (300 and 1000 mug/mL) inhibited neutrophil migration induced by piratoxin-I, whereas dermatan sulfate and chondroitin sulfate (30-1000 mug/mL each) had no effect. Heparitinase I and heparinase (300 mU/mL each) inhibited by 41.5 and 47%. respectively, piratoxin-l-induced chemotaxis, whereas heparitinase 11 and chondroitinase AC failed to affect the chemotaxis. the PAF receptor antagonist WEB 2086 (3-[4-(2-chlorophenyl)-9-methyl-6H-thienol-[3,2-f] [1,2,4]-triazolo-[4.3-a] [1,4]-diazepine-2-yl]-1-(4-morpholynil)-1-propionate) (0.1-10 muM) and the LTB4 synthesis inhibitor AA-861 [2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-1,4-benzoquinone] (0.1-10 muM) significantly inhibited the piratoxin-I-induced chemotaxis. Piratoxin-I (30-300 mug/mL) caused a concentration-dependent release of LTB4 Our results suggest that neutrophil migration in response to sPLA(2)s is independent of PLA activity, and involves an interaction of sPLA(2)s with cell surface heparin/heparan binding sites triggering the release of LTB4 and PAF. (C) 2002 Elsevier Science Inc. All rights reserved.
Biochemical Pharmacology. Oxford: Pergamon-Elsevier B.V., v. 63, n. 1, p. 65-72, 2002.