Abstract
Leukocyte de-activation in response to a mechanical stimulus may be an important mechanism to reduce inflammation in the circulation and cardiovascular complications. We examine here a specific form of leukocyte activation in the form of pseudopod projection, a process that is important during cell spreading and migration, but if it occurs in circulating leukocytes, may also lead to their entrapment in the microvascular network. Fresh neutrophils were activated with fMLP, suspended without adhesion to endothelium, and sheared in a cone-and-plate device while both shear stress and shear rate were measured. A fraction of the activated neutrophils retracted their pseudopods under the influence of fluid shear and returned to round shape. Pseudopod retraction was observed only in the presence of erythrocytes (at shear stresses up to ~25 dyn/cm2). At a constant hematocrit and increasing plasma viscosities with addition of macromolecules, the number of de-activated neutrophils scaled with shear stress and less so with shear rate. We examined a biochemical and rheological role of erythrocytes during shear de-activation of neutrophils. Addition of superoxide dismutase (SOD) in phosphate buffer served to enhance neutrophil de-activation by fluid shear. Replacement of erythrocytes by solid microspheres (5.4 μm) to simulate the particle properties of the erythrocytes, did not serve to enhance neutrophil de-activation unless in the presence of SOD. At higher shear stresses without erythrocytes (38–77 dyn/cm2), we also observed neutrophil de-activation but only in the presence of SOD. These results suggest that erythrocytes play an important role in neutrophil de-activation by reducing the superoxide level in plasma. Shear stress, rather than shear rate, is the key determinant that regulates neutrophil de-activation.
Similar content being viewed by others
References
Bao, X., C. Lu, and J. A. Frangos. Temporal gradient in shear but not steady shear stress induces PDGF-A and MCP-1 expression in endothelial cells: Role of NO, NF kappa B, and egr-1. Arterioscler. Thromb. Vasc. Biol. 9:996–1003, 1999.
Bengtsson, T., A. Fryden, S. Zalavary, P. A. Whiss, K. Orselius, and M. Grenegard. Platelets enhance neutrophil locomotion: Evidence for a role of P-selectin. Scand. J. Clin. Lab. Invest. 59:439–450, 1999.
Buerk, D. G., K. L. Lamkin-Kennard, and D. Jaron. Modeling the influence of superoxide dismutase on superoxide and nitric oxide interactions, including reversible inhibition oxygen consumption. Free Radic. Biol. Med. 34:1488–1503, 2003.
Buttrum, S. M., R. Haton, and G. B. Nash. Selectin-mediated rolling of neutrophils on immobilized platelets. Blood 82:1165–1174, 1993.
Chen, H.-Q., W. Tian, Y.-S. Chen, L. Li, J. Raum, and K.-L. P. Sung. Effect of steady and oscillatory shear stress on F-actin content and distribution in neutrophils. Biorheology 41:655–664, 2004.
Cinamon, G., V. Shinder, and R. Alon. Shear forces promote lymphocyte migration across vascular endothelium bearing apical chemokines. Nat. Immunol. 2:515–522, 2000.
Coughlin, M. F., and G. W. Schmid-Schönbein. Pseudopod projection and cell spreading of passive leukocytes in response to fluid shear stress. Biophys. J. 87:2035–2042, 2004.
Dewitz, T. S., T. C. Hung, R. R. Martin, and L. V. McIntire. Mechanical trauma in leukocytes. J. Lab. Clin. Med. 90:728–736, 1997.
Dong, C., R. Skalak, K.-L. P. Sung, G. W. Schmid-Schönbein, and S. Chien. Passive deformation analysis of human leukocytes. J. Biomech. Eng. 110:27–36, 1988.
Fukuda, S., T. Yasu, D. N. Predescu, and G. W. Schmid-Schönbein. Mechanisms for regulation of fluid shear stress response in circulating leukocytes. Circ. Res. 86:e13–e18, 2000.
Fukuda, S., and G. W. Schmid-Schönbein. Centrifugation attenuates the fluid shear response of circulating leukocytes. J. Leukoc. Biol. 72:133–139, 2002.
Fukuda, S., and G. W. Schmid-Schönbein. Regulation of CD18 expression on neutrophils in response to fluid shear stress. Proc. Natl. Acad. Sci. U.S.A. 100:13152–13157, 2003.
Fukuda, S., H. Mitsuoka, and G. W. Schmid-Schönbein. Leukocyte fluid shear response in the presence of glucocorticoid. J. Leukoc. Biol. 75:664–670, 2004.
Hampton, M. B., A. J. Kettle, and C. C. Winterbourn. Inside the neutrophil phagosome: Oxidants, myeloperoxidase, and bacterial killing. Blood 92:3007–3017, 1998.
Heikkila, R. E., F. S. Cabbat, and G. Cohen. In vivo inhibition of superoxide dismutase in mice by diethyldithiocarbamate. J. Biol. Chem. 251:2182–2185, 1976.
Hentzen, E., D. McDonough, L. McIntire, C. W. Smith, H. L Goldsmith, and S. I. Simon. Hydrodynamic shear and tethering through E-selectin signals phosphorylation of p38 MAP kinase and adhesion of human neutrophils. Ann. Biomed. Eng. 30:987–1001, 2002.
Hu, H., D. Varon, P. Hjemdahl, N. Savion, S. Schulman, and N. Li. Platelet–leukocyte aggregation under shear stress: Differential involvement of selectins and integrins. Thromb. Haemost. 90:679–687, 2003.
Johnson, R. M. Membrane stress increase cation permeability in red cells. Biophys. J. 67:1876–1881, 1994.
Kelner, M. J., and N. M. Alexander. Inhibition of erythrocyte superoxide dismutase by diethyldithiocarbamate also results in oxyhemoglobin-catalyzed glutathione depletion and methemoglobin production. J. Biol. Chem. 261:1636–1641, 1986.
Kitayama, J., A. Hidemura, H. Saito, and H. Nagawa. Shear stress affects migration behavior of polymorphonuclear cells arrested on endothelium. Cell. Immunol. 203:39–46, 2000.
Konstantopoulos, K., S. Neelamegham, A. R. Burns, E. Hentzen, G. S. Kansas, K. R. Snapp, E. L. Berg, J. D. Hellums, C. W. Smith, L. V. McIntire, and S. I. Simon. Venous levels of shear support neutrophil–platelet adhesion and neutrophil aggregation in blood via P-selectin and beta2-integrin. Circulation 98(9):873–882, 1998.
Konstantopoulos, K., S. Kukreti, and L. V. McIntire. Biomechanics of cell interactions in shear field. Adv. Drug Deliv. Rev. 33:141–164, 1998.
Kroll, M. H., J. D. Hellums, L. V. McIntire, A. I. Schafer, and J. L. Moake. Platelets and shear stress. Blood 88:1525–1541, 1996.
Lawrence, M. B., G. S. Kansas, E. J. Kunkel, and K. Ley. Threshold levels of fluid shear promote leukocyte adhesion through selectins (CD62L,P,E). J. Cell Biol. 136:717–727, 1997.
McAllister, T. N., and J. A. Frangos. Steady and transient fluid shear stress stimulate NO release in osteoblasts through distinct biochemical pathways. J. Bone Miner. Res. 14:930–936, 1999.
Moazzam, F., F. A. Delano, B. W. Zweifach, and G. W. Schmid-Schönbein. The leukocyte response to fluid stress. Proc. Natl. Acad. Sci. U.S.A. 94:5338–5343, 1997.
Nagata, K., T. Tsuji, N. Todoroki, Y. Katagiri, K. Tanoue, H. Yamazaki, N. Hanai, and T. Irimura. Activated platelets induce superoxide anion release by monocytes and neutrophils through P-selectin (CD62). J. Immunol. 151:3267–3273, 1993.
Nimeri, G., M. Majeed, H. Elwing, L. Ohman, J. Wettero, and T. Bengtsson. Oxygen radical production in neutrophils interacting with platelets and surface-immobilized plasma proteins: Role of tyrosin phosphorylation. J. Biomed. Mater. Res. 67A:439–447, 2003.
Rainger, G. E., C. D. Buckley, D. L. Simmons, and G. B. Nash. Neutrophils sense flow-generated stress and direct their migration through αv ß3-integrin. Am. J. Physiol. 276:H858–H864, 1999.
Ramos, C. L., M. J. Smith, S. K. Snapp, G. S. Kansas, G. W. Stickney, K. Ley, and M. B. Lawrence. Functional characterization of L-selectin ligands on human neutrophils and leukemia cell lines: Evidence for mucinlike ligand activity distinct from P-selectin glycoprotein ligand-1. Blood 91:1067–1075, 1998.
Resnick, N., H. Yahav, A. Shay-Salit, M. Shushy, S. Schubert, L. C. Zilberman, and E. Wofovitz. Fluid shear stress and the vascular endothelium: For better and for worse. Prog. Biophys. Mol. Biol. 81:177–199, 2003.
Ritter, L. S., D. S. Wilson, S. K. Williams, J. G. Copeland, and P. F. McDonagh. Early in reperfusion following myocardial ischemia, leukocyte activation is necessary for venular adhesion but not capillary retention. Microcirculation 2:315–327, 1995.
Schmid-Schönbein, G. W., E. B. Kistler, and T. E. Hugli. Mechanisms for cell activation and its consequences for biorheology and microcirculation: Multiorgan failure in shock. Biorheology 38:185–202, 2000.
Schmidtke, D. W., and S. L. Diamond. Direct observation of membrane tethers formed during neutrophil attachment to platelets or P-selectin under physiological flow. J. Cell Biol. 149:719–729, 2000.
Sugihara-Seki, M., and G. W. Schmid-Schönbein. The fluid shear stress distribution on the membrane of leukocytes in the microcirculation. J. Biomech. Eng. 125:628–638, 2003.
Sutton, D. W., and G. W. Schmid-Schönbein. Elevation of organ resistance due to leukocyte perfusion. Am. J. Physiol. 262:H1646–H1650, 1992.
Wettero, J., P. Tengvall, and T. Bengtsson. Platelets stimulated by IgG-coated surfaces bind and activate neutrophils through a selectin-dependent pathway. Biomaterials 24:1559–1573, 2003.
Worthen, G. S., B. Schwab, E. L. Elson, and G. P. Downey. Mechanics of stimulated neutrophils: Cell stiffening induces retention in capillaries. Science 245:183–186, 1989.
Zweifach, B. W., and H. H. Lipowsky. Pressure-flow relations in blood and lymph microcirculation. In: Handbook of Physiology, Section 2: The Cardiovascular System, edited by E. M. Renkin and C. C. Michel. Bethesda, MD: American Physiological Society, 1984, pp. 251–307.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Komai, Y., Schmid-Schönbein, G.W. De-Activation of Neutrophils in Suspension by Fluid Shear Stress: A Requirement for Erythrocytes. Ann Biomed Eng 33, 1375–1386 (2005). https://doi.org/10.1007/s10439-005-6768-6
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10439-005-6768-6