Force-dependent calcium signaling and its pathway of human neutrophils on P-selectin in flow

Abstract

P-selectin engagement of P-selectin glycoprotein ligand-1 (PSGL-1) causes circulating leukocytes to roll on and adhere to the vascular surface, and mediates intracellular calcium flux, a key but unclear event for subsequent arresting firmly at and migrating into the infection or injured tissue. Using a parallel plate flow chamber technique and intracellular calcium ion detector (Fluo-4 AM), the intracellular calciumflux of firmly adhered neutrophils on immobilized P-selectin in the absence of chemokines at various wall shear stresseswas investigated here in real time by fluorescence microscopy. The results demonstrated that P-selectin engagement of PSGL-1 induced the intracellular calcium flux of firmly adhered neutrophils in flow, increasing P-selectin concentration enhanced cellular calcium signaling, and, force triggered, enhanced and quickened the cytoplasmic calcium bursting of neutrophils on immobilized P-selectin. This P-selectin-induced calcium signaling should come from intracellular calcium release rather than extracellular calcium influx, and be along the mechano-chemical signal pathway involving the cytoskeleton, moesin and Spleen tyrosine kinase (Syk). These results provide a novel insight into the mechano-chemical regulation mechanism for P-selectininduced calcium signaling of neutrophils in flow.

Key words： neutrophils; P-selectin; calcium signaling; shear stress

References

1 Abbal C, Lambelet M, Bertaggia D, Gerbex C, Martinez M, Arcaro A, Schapira M, Spertini O (2006) Lipid raft adhesion receptors and Syk regulate selectin-dependent rolling under flow conditions. Blood 108:3352–3359

2 Barabé F, Paré G, Fernandes MJ, Bourgoin SG, Naccache PH (2002) Cholesterol-modulating agents selectively inhibit calcium influx induced by chemoattractants in human neutrophils. J Biol Chem 277:13473–13478

3 Clapham DE (1995) Calcium signaling. Cell 80:259–268

4 Davies MP, Hallam TJ, Merritt JE (1990) A role for calcium and protein kinase C in agonist-stimulated adhesion of human neutrophils. Biochem J 267:13–16

5 Falkenburger BH, Dickson EJ, Hille B (2013) Quantitative properties and receptor reserve of the DAG and PKC branch of Gq-coupled receptor signaling. J Gen Physiol 141:537–555

6 Geahlen RL (2014) Getting Syk: spleen tyrosine kinase as a therapeutic target. Trends Pharmacol Sci 35:414–422

7 Hogg N, Patzak I, Willenbrock F (2011) The insider’s guide to leukocyte integrin signalling and function. Nat Rev Immunol 11:416–426

8 Horejsi V, Hrdinka M (2014) Membrane microdomains in immunoreceptor signaling. FEBS Lett 588:2392–2397

9 Kannan KB, Barlos D, Hauser CJ (2007) Free cholesterol alters lipid raft structure and function regulating neutrophil Ca2+ entry and respiratory burst: correlations with calcium channel raft trafficking. J Immunol 178:5253–5261

10 Krishnan S, Juang YT, Chowdhury B, Magilavy A, Fisher CU, Nguyen H, Nambiar MP, Kyttaris V, Weinstein A, Bahjat R (2009) Differential expression and molecular associations of Syk in systemic lupus erythematosus Tcells. J Immunol 181:8145–8152

11 Kulathu Y, Hobeika E, Turchinovich G, Reth M (2008) The kinase Syk as an adaptor controlling sustained calcium signalling and B-cell development. EMBO J 27:1333–1344

12 Li Q, Wayman A, Lin J, Fang Y, Zhu C, Wu J (2016) Flow-enhanced stability of rolling adhesion through E-selectin. Biophys J 111:686–699

13 Ling Y, Zhang J, Li Q, Huang B, Fang Y (2013) Measuring site density of P-selectin on planar surface based on 125I labeling. BioTechnol 8:428–432

14 Ling Y, Fang Y, Yang X, Li Q, Lin Q, Wu J (2014) Regulation of shear stress on rolling behaviors of HL-60 cells on P-selectin. Sci China Phys Mech Astron 57:1998–2006

15 Liu B, Chen W, Evavold B, Zhu C (2014) Accumulation of dynamic catch bonds between TCR and agonist peptide-MHC triggers T cell signaling. Cell 157:357–368

16 Marois L, Paré G, Vaillancourt M, Rollet-Labelle E, Naccache PH (2011) Fc gamma RIIIb triggers raft-dependent calcium influx in IgG-mediated responses in human neutrophils. J Biol Chem 286:3509–3519

17 Marshall BT, Long M, Piper JW, Yago T, Mcever RP, Zhu C (2003) Direct observation of catch bonds involving cell-adhesion molecules. Nature 423:190–193

18 Matsumoto M, Hirata T (2016) Moesin regulates neutrophil rolling velocity in vivo. Cellular Immunol 304– 305:59–62

19 Mcever RP (2015) Selectins: initiators of leucocyte adhesion and signalling at the vascular wall. Cardiovasc Res 107:331–339

20 McEver RP, Cummings RD (1997) Role of PSGL-1 binding to selectins in leukocyte recruitment. J Clin Investig 100:97–103

21 Müller P, Langenbach A, Kaminski A, Rychly J (2013) Modulating the actin cytoskeleton affects mechanically induced signal transduction and differentiation in mesenchymal stem cells. PLoS ONE 8: e71283

22 Rebecchi MJ, Pentyala SN (2000) Structure, function, and control of phosphoinositide-specific phospholipase C. Physiol Rev 80:1291–1335

23 Romeo D, Zabucchi G, Miani N, Rossi F (1975) Ion movement across leukocyte plasma membrane and excitation of their metabolism. Nature 253:542–544

24 Schaff UY, Yamayoshi I, Tse T, Griffin D, Kibathi L, Simon SI (2008) Calcium flux in neutrophils synchronizes β 2 integrin adhesive and signaling events that guide inflammatory recruitment. Ann Biomed Eng 36:632–646

25 Serrador JM, Urzainqui A, Alonsolebrero JL, Cabrero JR, Montoya MC, Vicentemanzanares M, Yáñezmó M, Sánchezmadrid F (2002) A juxta-membrane amino acid sequence of P-selectin glycoprotein ligand-1 is involved in moesin binding and ezrin/ radixin/moesin-directed targeting at the trailing edge of migrating lymphocytes. Eur J Immunol 32:1560–1566

26 Shaffer MH, Dupree RS, Zhu P, Saotome I, Schmidt RF, Mcclatchey AI, Freedman BD, Burkhardt JK (2009) Ezrin and moesin function together to promote T cell activation. J Immunol 182:1021–1032

27 Shao B, Yago T, Setiadi H, Wang Y, Mehta-D’souza P, Fu J, Crocker PR, Rodgers W, Xia L, McEver RP (2015) O-glycans direct selectin ligands to lipid rafts on leukocytes. Proc Natl Acad Sci USA 112:8661–8666

28 Smolen JE, Korchak HM, Weissmann G (1981) The roles of extracellular and intracellular calcium in lysosomal enzyme release and superoxide anion generation by human neutrophils. Biochim Biophys Acta 677:512–520

29 Snapp KR, Heitzig CE, Kansas GS (2002) Attachment of the PSGL-1 cytoplasmic domain to the actin cytoskeleton is essential for leukocyte rolling on P-selectin. Blood 99:4494–4502

30 Stadtmann A, Brinkhaus L, Mueller H, Rossaint J, Bolomini-Vittori M, Bergmeier W, Van Aken H, Wagner DD, Laudanna C, Ley K (2011) Rap1a activation by CalDAG-GEFI and p38 MAPK is involved in E-selectin-dependent slow leukocyte rolling. Eur J Immunol 41:2074–2085

31 Takata M, Sabe H, Hata A, Inazu T, Homma Y, Nukada T, Yamamura H, Kurosaki T (1994) Tyrosine kinases Lyn and Syk regulate B cell receptor-coupled Ca2+ mobilization through distinct pathways. EMBO J 13:1341–1349

32 Urzainqui A, Serrador JM, Viedma F, Yáñez-Mó MA, Rodríguez A, Corbı́ AL, Alonso-Lebrero JL, Luque A, Deckert M, Vázquez J (2002) ITAM-based interaction of ERM proteins with Syk mediates signaling by the leukocyte adhesion receptor PSGL-1. Immunity 17:401–412

33 Wang N, Butler JP, Ingber DE (1993) Mechanotransduction across the cell surface and through the cytoskeleton. Science 260:1124–1127

34 Wang XG, Cheng YP, Ba XQ (2006) Engagement of PSGL-1 enhances beta(2)-integrin-involved adhesion of neutrophils to recombinant ICAM-1. Acta Pharmacol Sin 27:617–622

35 Yago T, Shao B, Miner JJ, Yao L, Klopocki AG, Maeda K, Coggeshall KM, McEver RP(2010) E-selectin engages PSGL-1 and CD44 through a common signaling pathway to induce integrin αLβ2-mediated slow leukocyte rolling. Blood 116:485–494

36 Yin HL, Stossel TP (1979) Control of cytoplasmic actin gel-sol transformation by gelsolin, a calcium-dependent regulatory protein. Nature 281:583–586

37 Yuan SY, Shen Q, Rigor RR, Wu MH (2012) Neutrophil transmigration, focal adhesion kinase and endothelial barrier function. Microvasc Res 83:82–88

38 Zarbock A, Ley K (2009) Neutrophil Adhesion and Activation under Flow. Microcirculation 16:31–42