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Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro

BACKGROUND: Cytosolic gIVaPLA(2 )is a critical enzyme in the generation of arachidonate metabolites and in induction of β(2)-integrin adhesion in granulocytes. We hypothesized that gIVaPLA(2 )activation also is an essential downstream step for post adhesive migration of PMN in vitro. METHODS: Migrat...

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Autores principales: Meliton, Angelo Y, Muñoz, Nilda M, Meliton, Lucille N, Binder, David C, Osan, Christopher M, Zhu, Xiangdong, Dudek, Steven M, Leff, Alan R
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2848033/
https://www.ncbi.nlm.nih.gov/pubmed/20298597
http://dx.doi.org/10.1186/1476-9255-7-14
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author Meliton, Angelo Y
Muñoz, Nilda M
Meliton, Lucille N
Binder, David C
Osan, Christopher M
Zhu, Xiangdong
Dudek, Steven M
Leff, Alan R
author_facet Meliton, Angelo Y
Muñoz, Nilda M
Meliton, Lucille N
Binder, David C
Osan, Christopher M
Zhu, Xiangdong
Dudek, Steven M
Leff, Alan R
author_sort Meliton, Angelo Y
collection PubMed
description BACKGROUND: Cytosolic gIVaPLA(2 )is a critical enzyme in the generation of arachidonate metabolites and in induction of β(2)-integrin adhesion in granulocytes. We hypothesized that gIVaPLA(2 )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 gIVaPLA(2), 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. GIVaPLA(2 )enzyme activity was analyzed using [(14)C-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 gIVaPLA(2 )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 gIVaPLA(2 )corresponded to the magnitude of blockade of PMN migration. Neither AA nor LTB(4 )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 gIVaPLA(2 )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 gIVaPLA(2 )to the nucleus, b) change in cell shape, c) polymerization of F-actin, and d) chemoattractant/migration of PMN in vitro. Inhibition of gIVaPLA(2 )blocks the deformability and subsequent migration of PMNs caused by IL-8/CXCL8. Our data suggest that activation of gIVaPLA(2 )is an essential step in PMN migration in vitro.
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spelling pubmed-28480332010-04-01 Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro Meliton, Angelo Y Muñoz, Nilda M Meliton, Lucille N Binder, David C Osan, Christopher M Zhu, Xiangdong Dudek, Steven M Leff, Alan R J Inflamm (Lond) Research BACKGROUND: Cytosolic gIVaPLA(2 )is a critical enzyme in the generation of arachidonate metabolites and in induction of β(2)-integrin adhesion in granulocytes. We hypothesized that gIVaPLA(2 )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 gIVaPLA(2), 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. GIVaPLA(2 )enzyme activity was analyzed using [(14)C-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 gIVaPLA(2 )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 gIVaPLA(2 )corresponded to the magnitude of blockade of PMN migration. Neither AA nor LTB(4 )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 gIVaPLA(2 )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 gIVaPLA(2 )to the nucleus, b) change in cell shape, c) polymerization of F-actin, and d) chemoattractant/migration of PMN in vitro. Inhibition of gIVaPLA(2 )blocks the deformability and subsequent migration of PMNs caused by IL-8/CXCL8. Our data suggest that activation of gIVaPLA(2 )is an essential step in PMN migration in vitro. BioMed Central 2010-03-18 /pmc/articles/PMC2848033/ /pubmed/20298597 http://dx.doi.org/10.1186/1476-9255-7-14 Text en Copyright ©2010 Meliton et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Meliton, Angelo Y
Muñoz, Nilda M
Meliton, Lucille N
Binder, David C
Osan, Christopher M
Zhu, Xiangdong
Dudek, Steven M
Leff, Alan R
Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title_full Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title_fullStr Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title_full_unstemmed Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title_short Cytosolic group IVa phospholipase A(2 )mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro
title_sort cytosolic group iva phospholipase a(2 )mediates il-8/cxcl8-induced transmigration of human polymorphonuclear leukocytes in vitro
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2848033/
https://www.ncbi.nlm.nih.gov/pubmed/20298597
http://dx.doi.org/10.1186/1476-9255-7-14
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