fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils
N-Formyl-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) induce similar intracellular signalling profiles; but only fMLP induces interleukin-8 (IL-8) release and nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase activity in neutrophils. Because the role o...
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2015/120348 |
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| author | María A. Hidalgo María D. Carretta Stefanie E. Teuber Cristian Zárate Leonardo Cárcamo Ilona I. Concha Rafael A. Burgos |
| author_facet | María A. Hidalgo María D. Carretta Stefanie E. Teuber Cristian Zárate Leonardo Cárcamo Ilona I. Concha Rafael A. Burgos |
| author_sort | María A. Hidalgo |
| collection | DOAJ |
| description | N-Formyl-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) induce similar intracellular signalling profiles; but only fMLP induces interleukin-8 (IL-8) release and nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase activity in neutrophils. Because the role of ROS on IL-8 release in neutrophils is until now controversial, we assessed if NADPH oxidase is involved in the IL-8 secretions and PI3K/Akt, MAPK, and NF-κB pathways activity induced by fMLP. Neutrophils were obtained from healthy volunteers. IL-8 was measured by ELISA, IL-8 mRNA by qPCR, and ROS production by luminol-amplified chemiluminescence, reduction of ferricytochrome c, and FACS. Intracellular pH changes were detected by spectrofluorescence. ERK1/2, p38 MAPK, and Akt phosphorylation were analysed by immunoblotting and NF-κB was analysed by immunocytochemistry. Hydroxy-3-methoxyaceto-phenone (HMAP), diphenyleneiodonium (DPI), and siRNA Nox2 reduced the ROS and IL-8 release in neutrophils treated with fMLP. HMAP, DPI, and amiloride (a Na+/H+ exchanger inhibitor) inhibited the Akt phosphorylation and did not affect the p38 MAPK and ERK1/2 activity. DPI and HMAP reduced NF-κB translocation induced by fMLP. We showed that IL-8 release induced by fMLP is dependent on NADPH oxidase, and ROS could play a redundant role in cell signalling, ultimately activating the PI3K/Akt and NF-κB pathways in neutrophils. |
| format | Article |
| id | doaj-art-02a7784e20a546dd83ce3692d45bbbbb |
| institution | Kabale University |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-02a7784e20a546dd83ce3692d45bbbbb2025-02-03T01:23:50ZengWileyJournal of Immunology Research2314-88612314-71562015-01-01201510.1155/2015/120348120348fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human NeutrophilsMaría A. Hidalgo0María D. Carretta1Stefanie E. Teuber2Cristian Zárate3Leonardo Cárcamo4Ilona I. Concha5Rafael A. Burgos6Laboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileLaboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileLaboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileLaboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileLaboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileInstitute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileLaboratory of Molecular Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Independencia 631, 5110566 Valdivia, ChileN-Formyl-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) induce similar intracellular signalling profiles; but only fMLP induces interleukin-8 (IL-8) release and nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase activity in neutrophils. Because the role of ROS on IL-8 release in neutrophils is until now controversial, we assessed if NADPH oxidase is involved in the IL-8 secretions and PI3K/Akt, MAPK, and NF-κB pathways activity induced by fMLP. Neutrophils were obtained from healthy volunteers. IL-8 was measured by ELISA, IL-8 mRNA by qPCR, and ROS production by luminol-amplified chemiluminescence, reduction of ferricytochrome c, and FACS. Intracellular pH changes were detected by spectrofluorescence. ERK1/2, p38 MAPK, and Akt phosphorylation were analysed by immunoblotting and NF-κB was analysed by immunocytochemistry. Hydroxy-3-methoxyaceto-phenone (HMAP), diphenyleneiodonium (DPI), and siRNA Nox2 reduced the ROS and IL-8 release in neutrophils treated with fMLP. HMAP, DPI, and amiloride (a Na+/H+ exchanger inhibitor) inhibited the Akt phosphorylation and did not affect the p38 MAPK and ERK1/2 activity. DPI and HMAP reduced NF-κB translocation induced by fMLP. We showed that IL-8 release induced by fMLP is dependent on NADPH oxidase, and ROS could play a redundant role in cell signalling, ultimately activating the PI3K/Akt and NF-κB pathways in neutrophils.http://dx.doi.org/10.1155/2015/120348 |
| spellingShingle | María A. Hidalgo María D. Carretta Stefanie E. Teuber Cristian Zárate Leonardo Cárcamo Ilona I. Concha Rafael A. Burgos fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils Journal of Immunology Research |
| title | fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils |
| title_full | fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils |
| title_fullStr | fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils |
| title_full_unstemmed | fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils |
| title_short | fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils |
| title_sort | fmlp induced il 8 release is dependent on nadph oxidase in human neutrophils |
| url | http://dx.doi.org/10.1155/2015/120348 |
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