Background/Aims: This study investigated the priming effect of sphingosine 1-phosphate (S1P) on formyl-Met-Leu-Phe-OH (fMLP)-activated neutrophils, by specific analysis of the neutrophil respiratory burst and the signaling pathway involved in S1P activity. Methods: The neutrophil respiratory burst was indirectly detected by the cytochrome c reduction method and the dihydrorhodamine 123 staining method. The signal transduction pathways of neutrophil respiratory burst primed by S1P were detected by Western blotting. Results: Our results showed that the S1P receptors (S1PRs) 1, 4 and 5 were the predominantly expressed neutrophil S1PRs at the cDNA level. After pretreatment with S1P, the fMLP-activated neutrophils released increased levels of superoxide anions. PI3K and Akt proteins were involved in the signaling pathway of the neutrophil respiratory burst primed by S1P. Conclusion: The results indicate that S1P is a new priming reagent for neutrophils and primes the respiratory burst of fMLP-activated neutrophils. S1P interacts with its receptors on neutrophils, resulting in NADPH oxidase activation by the PI3K-Akt cell signaling pathway and induction of the neutrophil respiratory burst.

Keywords:
Neutrophils, Respiratory burst, Sphingosine 1-phosphate, Superoxide anion, Phosphoinositide 3-kinase
1.
Balloy V, Chignard M: The innate immune response to Aspergillus fumigatus. Microbes Infect 2009;11:919-927.
2.
Holmes B, Page AR, Good RA: Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J Clin Invest 1967;46:1422-1432.
3.
Segal BH, Romani LR: Invasive aspergillosis in chronic granulomatous disease. Med Mycol 2009;47:S282-S290.
4.
Bode C, Sensken SC, Peest U, Beutel G, Thol F, Levkau B, Li Z, Bittman R, Huang T, Tölle M, van der Giet M, Gräler MH: Erythrocytes serve as a reservoir for cellular and extracellular sphingosine 1-phosphate. J Cell Biochem 2010;109:1232-1243.
5.
Rosen H, Gonzalez-Cabrera PJ, Sanna MG, Browm S: Sphingosine 1-phosphate receptor signaling. Annu Rev Biochem 2009;78:743-768.
6.
Pappu R, Schwab SR, Cornelissen I, Pereira JP, Regard JB, Xu Y, Camerer E, Zheng YW, Huang Y, Cyster JG, Coughlin SR: Promotion of lymphocyte egress into blood and lymph by distinct sources of sphingosine-1-phosphate. Science 2007;316:295-298.
7.
Zachariah MA, Cyster JG: Neural crest-derived pericytes promote egress of mature thymocytes at the corticomedullary junction. Science 2010;328:1129-1135.
8.
Pham TH, Baluk P, Xu Y, Grigorova I, Bankovich AJ, Pappu R, Coughlin SR, McDonald DM, Schwab SR, Cyster JG: Lymphatic endothelial cell sphingosine kinase activity is required for lymphocyte egress and lymphatic patterning. J Exp Med 2010;207:17-27.
9.
Chi H: Sphingosine-1-phosphate and immune regulation: trafficking and beyond. Trends Pharmacol Sci 2011;32:16-24.
10.
Lister KJ, James WG, Hickey MJ: Immune complexes mediate rapid alterations in microvascular permeability: roles for neutrophils, complement, and platelets. Microcirculation 2007;14:709-722.
11.
Ancellin N, Colmont C, Su J, Li Q, Mittereder N, Chae SS, Stefansson S, Liau G, Hla T: Extracellular export of sphingosine kinase-1 enzyme: sphingosine 1-phosphate generation and the induction of angiogenic vascular maturation. J Biol Chem 2002;277:6667-6675.
12.
Florey O, Haskard DO: Sphingosine 1-phosphate enhances Fcgamma receptor-mediated neutrophil activation and recruitment under flow conditions. J Immunol 2009;183:2330-2336.
13.
Tsuboi N, Asano K, Lauterbach M, Mayadas TN: Human neutrophil Fcgamma receptors initiate and play specialized nonredundant roles in antibody-mediated inflammatory diseases. Immunity 2008;28:833-846.
14.
Gao XP, Zhu X, Fu J, Liu Q, Frey RS, Malik AB: Blockade of class IA phosphoinositide 3-kinase in neutrophils prevents NADPH oxidase activation- and adhesion-dependent inflammation. J Biol Chem 2007;282:6116-6125.
15.
Frey RS, Gao X, Javaid K, Siddiqui SS, Rahman A, Malik AB: Phosphatidylinositol 3-kinase gamma signaling through protein kinase Czeta induces NADPH oxidase-mediated oxidant generation and NF-kappaB activation in endothelial cells. J Biol Chem 2006;281:16128-16138.
16.
Boulven I, Levasseur S, Marois S, Paré G, Rollet-Labelle E, Naccache PH: Class IA phosphatidylinositide 3-kinases, rather than p110 gamma, regulate formyl-methionyl-leucyl-phenylalanine-stimulated chemotaxis and superoxide production in differentiated neutrophil-like PLB-985 cells. J Immunol 2006;176:7621-7627.
17.
Bitzinger DI, Schlachetzki F, Lindner R, Trabold B, Dittmar MS: Flow-cytometry measurement of respiratory burst in rat polymorphonuclear granulocytes: comparison of four cell preparation procedures, and concentration-response evaluation of soluble stimulants. Cytometry A 2008;73:643-650.
18.
Dahlgren C, Karlsson A: Respiratory burst in human neutrophils. J Immunol Methods 1999;232:3-14.
19.
Paape MJ, Bannerman DD, Zhao X, Lee JW: The bovine neutrophil: structure and function in blood and milk. Vet Res 2003;34:597-627.
20.
Cardo LJ, Wilder D, Salata J: Neutrophil priming, caused by cell membranes and microvesicles in packed red blood cell units, is abrogated by leukocyte depletion at collection. Transfus Apher Sci 2008;38:117-125.
21.
Liu G, Burns S, Huang G, Boyd K, Proia RL, Flavell RA, Chi H: The receptor S1P1 overrides regulatory T cell-mediated immune suppression through Akt-mTOR. Nat Immunol 2009;10:769-777.
22.
Müller H, Hofer S, Kaneider N, Neuwirt H, Mosheimer B, Mayer G, Konwalinka G, Heufler C, Tiefenthaler M: The immunomodulator FTY720 interferes with effector functions of human monocyte-derived dendritic cells. Eur J Immunol 2005;35:533-545.
23.
Rahaman M, Costello RW, Belmonte KE, Gendy SS, Walsh MT: Neutrophil sphingosine 1-phosphate and lysophosphatidic acid receptors in pneumonia. Am J Respir Cell Mol Biol 2006;34:233-241.
24.
Boyle KB, Gyori D, Sindrilaru A, Scharffetter-Kochanek K, Taylor PR, Mócsai A, Stephens LR, Hawkins PT: Class IA phosphoinositide 3-kinase β and δ regulate neutrophil oxidase activation in response to Aspergillus fumigatus hyphae. J Immunol 2011;186:2978-2989.
© 2015 S. Karger AG, Basel
2015
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.