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Journal of Inflammation
Published in: Journal of Inflammation 1/2010

Open Access 01-12-2010 | Research

Cytosolic group IVa phospholipase A2 mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro

Authors: Angelo Y Meliton, Nilda M Muñoz, Lucille N Meliton, David C Binder, Christopher M Osan, Xiangdong Zhu, Steven M Dudek, Alan R Leff

Published in: Journal of Inflammation | Issue 1/2010

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Abstract

Background

Cytosolic gIVaPLA2 is a critical enzyme in the generation of arachidonate metabolites and in induction of β2-integrin adhesion in granulocytes. We hypothesized that gIVaPLA2 activation also is an essential downstream step for post adhesive migration of PMN in vitro.

Methods

Migration of PMNs caused by IL-8/CXCL8 was assessed using a transwell migration chamber. PMNs were pretreated with two structurally unrelated inhibitors of gIVaPLA2, arachidonyl trifluoromethylketone (TFMK) or pyrrophenone, prior to IL-8/CXCL8 exposure. The fraction of migrated PMNs present in the lower chamber was measured as total myeloperoxidase content. GIVaPLA2 enzyme activity was analyzed using [14C-PAPC] as specific substrate F-actin polymerization and cell structure were examined after rhodamine-phalloidin staining.

Results

IL-8/CXCL8-induced migration of PMNs was elicited in concentration- and time-dependent manner. Time-related phosphorylation and translocation of cytosolic gIVaPLA2 to the nucleus was observed for PMNs stimulated with IL-8/CXCL8 in concentration sufficient to cause upstream phosphorylation of MAPKs (ERK-1/2 and p38) and Akt/PKB. Inhibition of gIVaPLA2 corresponded to the magnitude of blockade of PMN migration. Neither AA nor LTB4 secretion was elicited following IL-8/CXCL8 activation. In unstimulated PMNs, F-actin was located diffusely in the cytosol; however, a clear polarized morphology with F-actin-rich ruffles around the edges of the cell was observed after activation with IL-8/CXCL8. Inhibition of gIVaPLA2 blocked change in cell shape and migration caused by IL-8/CXCL8 but did not cause F-actin polymerization or translocation of cytosolic F-actin to inner leaflet of the PMN membrane.

Conclusion

We demonstrate that IL-8/CXCL8 causes a) phosphorylation and translocation of cytosolic gIVaPLA2 to the nucleus, b) change in cell shape, c) polymerization of F-actin, and d) chemoattractant/migration of PMN in vitro. Inhibition of gIVaPLA2 blocks the deformability and subsequent migration of PMNs caused by IL-8/CXCL8. Our data suggest that activation of gIVaPLA2 is an essential step in PMN migration in vitro.
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Literature
1.
Sai J, Raman D, Liu Y, Wikswo J, Richmond A: Parallel phosphatidylinositol 3-kinase (PI3K)-dependent and Src-dependent pathways lead to CXCL8-mediated Rac2 activation and chemotaxis. J Biol Chem. 2008, 283: 26538-26547. 10.1074/jbc.M805611200.PubMedPubMedCentralCrossRef
2.
Lane HC, Anand AR, Ganju RK: Cbl and Akt regulate CXCL8-induced and CXCR1- and CXCR2-mediated chemotaxis. Int Immunol. 2006, 18: 1315-1325. 10.1093/intimm/dxl064.PubMedCrossRef
3.
Qian Y, Corum L, Meng Q, Blenis J, Zheng JZ, Shi X, Flynn DC, Jiang BH: PI3K induced actin filament remodeling through Akt and p70S6K1: implication of essential role in cell migration. Am J Physiol Cell Physiol. 2004, 286: C153-163. 10.1152/ajpcell.00142.2003.PubMedCrossRef
4.
Cicchetti G, Allen PG, Glogauer M: Chemotactic signaling pathways in neutrophils: from receptor to actin assembly. Crit Rev Oral Biol Med. 2002, 13: 220-228. 10.1177/154411130201300302.PubMedCrossRef
5.
Katanaev VL: Signal transduction in neutrophil chemotaxis. Biochemistry (Mosc). 2001, 66: 351-368. 10.1023/A:1010293809553.CrossRef
6.
Niggli V: Signaling to migration in neutrophils: importance of localized pathways. Int J Biochem Cell Biol. 2003, 35: 1619-1638. 10.1016/S1357-2725(03)00144-4.PubMedCrossRef
7.
Kobashi C, Asamizu S, Ishiki M, Iwata M, Usui I, Yamazaki K, Tobe K, Kobayashi M, Urakaze M: Inhibitory effect of IL-8 on insulin action in human adipocytes via MAP kinase pathway. J Inflamm (Lond). 2009, 6: 25-10.1186/1476-9255-6-25.CrossRef
8.
Schaloske RH, Dennis EA: The phospholipase A2 superfamily and its group numbering system. Biochim Biophys Acta. 2006, 1761: 1246-1259.PubMedCrossRef
9.
Dennis EA: The growing phospholipase A2 superfamily of signal transduction enzymes. Trends Biochem Sci. 1997, 22: 1-2. 10.1016/S0968-0004(96)20031-3.PubMedCrossRef
10.
Bonventre JV, Huang Z, Taheri MR, O'Leary E, Li E, Moskowitz MA, Sapirstein A: Reduced fertility and postischaemic brain injury in mice deficient in cytosolic phospholipase A2. Nature. 1997, 390: 622-625. 10.1038/37635.PubMedCrossRef
11.
Burke JE, Dennis EA: Phospholipase A2 structure/function, mechanism, and signaling. J Lipid Res. 2009, 50 (Suppl): S237-242. 10.1194/jlr.R800033-JLR200.PubMedPubMedCentral
12.
Valentin E, Lambeau G: Increasing molecular diversity of secreted phospholipases A2 and their receptors and binding proteins. Biochim Biophys Acta. 2000, 1488: 59-70.PubMedCrossRef
13.
Murakami M, Kudo I: Diversity and regulatory functions of mammalian secretory phospholipase A2s. Adv Immunol. 2001, 77: 163-194. full_text.PubMedCrossRef
14.
Leslie CC: Properties and regulation of cytosolic phospholipase A2. J Biol Chem. 1997, 272: 16709-16712. 10.1074/jbc.272.27.16709.PubMedCrossRef
15.
Balsinde J, Dennis EA: Function and inhibition of intracellular calcium-independent phospholipase A2. J Biol Chem. 1997, 272: 16069-16072. 10.1074/jbc.272.26.16069.PubMedCrossRef
16.
Balsinde J, Perez R, Balboa MA: Calcium-independent phospholipase A2 and apoptosis. Biochim Biophys Acta. 2006, 1761: 1344-1350.PubMedCrossRef
17.
Stafforini DM: Biology of platelet-activating factor acetylhydrolase (PAF-AH, lipoprotein associated phospholipase A2). Cardiovasc Drugs Ther. 2009, 23: 73-83. 10.1007/s10557-008-6133-8.PubMedCrossRef
18.
Arai H, Koizumi H, Aoki J, Inoue K: Platelet-activating factor acetylhydrolase (PAF-AH). J Biochem. 2002, 131: 635-640.PubMedCrossRef
19.
Abe A, Hiraoka M, Shayman JA: A role for lysosomal phospholipase A2 in drug induced phospholipidosis. Drug Metab Lett. 2007, 1: 49-53. 10.2174/187231207779814292.PubMedCrossRef
20.
Niknami M, Patel M, Witting PK, Dong Q: Molecules in focus: cytosolic phospholipase A2-alpha. Int J Biochem Cell Biol. 2009, 41: 994-997. 10.1016/j.biocel.2008.07.017.PubMedCrossRef
21.
Zhu X, Munoz NM, Kim KP, Sano H, Cho W, Leff AR: Cytosolic phospholipase A2 activation is essential for β1 and β2 integrin-dependent adhesion of human eosinophils. J Immunol. 1999, 163: 3423-3429.PubMed
22.
Sano H, Zhu X, Sano A, Boetticher EE, Shioya T, Jacobs B, Munoz NM, Leff AR: Extracellular signal-regulated kinase 1/2-mediated phosphorylation of cytosolic phospholipase A2 is essential for human eosinophil adhesion to fibronectin. J Immunol. 2001, 166: 3515-3521.PubMedCrossRef
23.
Zhu X, Jacobs B, Boetticher E, Myou S, Meliton A, Sano H, Lambertino AT, Munoz NM, Leff AR: IL-5-induced integrin adhesion of human eosinophils caused by ERK1/2-mediated activation of cPLA2. J Leukoc Biol. 2002, 72: 1046-1053.PubMed
24.
Zhu X, Sano H, Kim KP, Sano A, Boetticher E, Munoz NM, Cho W, Leff AR: Role of mitogen-activated protein kinase-mediated cytosolic phospholipase A2 activation in arachidonic acid metabolism in human eosinophils. J Immunol. 2001, 167: 461-468.PubMedCrossRef
25.
Myou S, Leff AR, Myo S, Boetticher E, Meliton AY, Lambertino AT, Liu J, Xu C, Munoz NM, Zhu X: Activation of group IV cytosolic phospholipase A2 in human eosinophils by phosphoinositide 3-kinase through a mitogen-activated protein kinase-independent pathway. J Immunol. 2003, 171: 4399-4405.PubMedCrossRef
26.
Meliton AY, Munoz NM, Osan CM, Meliton LN, Leff AR: Leukotriene D4 activates β2-integrin adhesion in human polymorphonuclear leukocytes. Eur Respir J. 2009,
27.
Meliton AY, Munoz NM, Zhu X, Leff AR: Attenuated translocation of group IVa phospholipase A2 and up-regulated annexin-1 synthesis by glucocorticoid blocks β2-integrin adhesion in neutrophils. J Leukoc Biol. 2008, 83: 344-351. 10.1189/jlb.0707495.PubMedCrossRef
28.
Nolan S, Dixon R, Norman K, Hellewell P, Ridger V: Nitric oxide regulates neutrophil migration through microparticle formation. Am J Pathol. 2008, 172: 265-273. 10.2353/ajpath.2008.070069.PubMedPubMedCentralCrossRef
29.
Sano A, Zhu X, Sano H, Munoz NM, Boetticher E, Leff AR: Regulation of eosinophil function by phosphatidylinositol-specific PLC and cytosolic PLA2. Am J Physiol Lung Cell Mol Physiol. 2001, 281: L844-851.PubMed
30.
Casas J, Valdearcos M, Pindado J, Balsinde J, Balboa MA: The cationic cluster of group IVA phospholipase A2 (Lys488/Lys541/Lys543/Lys544) is involved in translocation of the enzyme to phagosomes in human macrophages. J Lipid Res. 2009, 51 (2): 388-99. 10.1194/jlr.M001461.PubMedCrossRef
31.
Meliton AY, Munoz NM, Liu J, Lambertino AT, Boetticher E, Myo S, Myou S, Zhu X, Johnson M, Leff AR: Blockade of LTC4 synthesis caused by additive inhibition of gIV-PLA2 phosphorylation: Effect of salmeterol and PDE4 inhibition in human eosinophils. J Allergy Clin Immunol. 2003, 112: 404-410. 10.1067/mai.2003.1637.PubMedCrossRef
32.
Cooper JA, Loftus DJ, Frieden C, Bryan J, Elson EL: Localization and mobility of gelsolin in cells. J Cell Biol. 1988, 106: 1229-1240. 10.1083/jcb.106.4.1229.PubMedCrossRef
33.
Sano A, Munoz NM, Sano H, Choi J, Zhu X, Jacobs B, Leff AR: Inhibition of cPLA2 translocation and leukotriene C4 secretion by fluticasone propionate in exogenously activated human eosinophils. Am J Respir Crit Care Med. 1999, 159: 1903-1909.PubMedCrossRef
34.
Szczur K, Zheng Y, Filippi MD: The small Rho GTPase Cdc42 regulates neutrophil polarity via CD11b integrin signaling. Blood. 2009
35.
Inoue T, Meyer T: Synthetic activation of endogenous PI3K and Rac identifies an AND-gate switch for cell polarization and migration. PLoS One. 2008, 3: e3068-10.1371/journal.pone.0003068.PubMedPubMedCentralCrossRef
36.
Fujimura M, Xiu Q, Tsujiura M, Tachibana H, Myou S, Matsuda T, Matsushima K: Role of leukotriene B4 in bronchial hyperresponsiveness induced by interleukin-8. Eur Respir J. 1998, 11: 306-311. 10.1183/09031936.98.11020306.PubMedCrossRef
37.
Saito H, Minamiya Y, Saito S, Ogawa J: Endothelial Rho and Rho kinase regulate neutrophil migration via endothelial myosin light chain phosphorylation. J Leukoc Biol. 2002, 72: 829-836.PubMed
38.
Snapper SB, Meelu P, Nguyen D, Stockton BM, Bozza P, Alt FW, Rosen FS, von Andrian UH, Klein C: WASP deficiency leads to global defects of directed leukocyte migration in vitro and in vivo. J Leukoc Biol. 2005, 77: 993-998. 10.1189/jlb.0804444.PubMedCrossRef
39.
Blake KM, Carrigan SO, Issekutz AC, Stadnyk AW: Neutrophils migrate across intestinal epithelium using β2 integrin (CD11b/CD18)-independent mechanisms. Clin Exp Immunol. 2004, 136: 262-268. 10.1111/j.1365-2249.2004.02429.x.PubMedPubMedCentralCrossRef
Metadata
Title
Cytosolic group IVa phospholipase A2 mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
Authors
Angelo Y Meliton
Nilda M Muñoz
Lucille N Meliton
David C Binder
Christopher M Osan
Xiangdong Zhu
Steven M Dudek
Alan R Leff
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Journal of Inflammation / Issue 1/2010
Electronic ISSN: 1476-9255
DOI
https://doi.org/10.1186/1476-9255-7-14

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