Advertisement

A Polymer Mixing Approach to Chemoattractant-Stimulated Neutrophil Adhesion

  • Donald J. L. McIver
  • Samuel Schürch

Abstract

We have used photometric and aqueous phase polymer affinity techniques to investigate the kinetics and surface thermodynamics of neutrophilic leukocyte responses to the soluble bacterial chemotaxin N-formyl methionyl leucyl phenylalanine (fMLP). Unlike the relatively slow responses of macrophages to particles, neutrophils respond to chemoattractants within seconds and display a complex series of time-dependent changes which in part reflect cellular adaptation to the chemotaxin by surface receptor internalisation [1,2]. The kinetics of these changes can be resolved by photometric methods, but not by the relatively slow techniques of macroscopic polymer phase wetting. We have therefore used the fungal metabolite cytochalasin B to prevent reversal of the rapid neutrophil responses to fMLP. Under these conditions, wetting of the neutrophils by dextran/PEG phases is essentially independent of time, and depends only on the dose of chemoattractant. The dose-response curves for fMLP induced cell aggregation and increased dextran phase affinity are similar, each with an ED50 of 2 x 10−8 M, suggesting that the two processes may have the same basis, for example in specific granule glycoproteins which become expressed on the cell surface during aggregation. By contrast, chemotactic cell surface ruffling (assessed either by digital image analysis of photomicrographs or by perpendicular light scattering from the cell suspensions) and oxygen radical-dependent chemiluminescence have dose-response curves with values of ED50 ten times lower and higher, respectively, than aggregation and phase affinity changes.

Keywords

Contact Angle Light Transmission Interfacial Free Energy Surface Thermodynamic Cell BioI 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    S.T. Hoffstein, R.S. Friedman and G. Weissman, Degranulation, membrane addition and shape change during chemotactic factor-induced aggregation of human neutrophils, J. Cell Biol. 95:234 (1982)PubMedCrossRefGoogle Scholar
  2. 2.
    I. Yuli and R. Snyderman, Rapid changes in light scattering from human polymorphonuclear leukocytes exposed to chemoattractants. Discrete responses correlated with chemotactic and secretory functions, J. Clin. Invest. 73:1408 (1984)PubMedCrossRefGoogle Scholar
  3. 3.
    E.A. Evans and R. Skalak, “Mechanics and Thermodynamics of Biomembranes,” CRC Press, Boca Raton, Fl. (1980)Google Scholar
  4. 4.
    I. Yuli and R. Snyderman, Light scattering by polymorphonuclear leukocytes stimulated to aggregate under various pharmacological conditions, Blood 64:649 (1984)PubMedGoogle Scholar
  5. 5.
    P. Latimer, Light scattering and absorption as methods of studying cell population parameters, Ann. Rev. Biophys. Bioeng. 11:129 (1982)CrossRefGoogle Scholar
  6. 6.
    L.A. Sklar, G.M. Omann and R.G. Painter, Relationship of actin polymerisation and depolymerisation to light scattering in human neutrophils: dependence on receptor occupancy and intracellular Ca++, J. Cell Biol. 101:1161 (1985)PubMedCrossRefGoogle Scholar
  7. 7.
    D.J.L. McIver and S. Schürch, Polymer mixing and the thermodynamics of cell adhesion at fluid interfaces, J. Adhesion. 22:253 (1987)CrossRefGoogle Scholar
  8. 8.
    S. Schürch and D.J.L. McIver, Interfacial free energies at lipid-water interfaces. Comparison of equation-of-state predictions with direct experimental measurements, J. Colloid Interface Sci. 83:301 (1981)CrossRefGoogle Scholar
  9. 9.
    S. Schürch, D.F. Gerson and D.J.L. McIver, Determination of cell-medium interfacial tensions from contact angles in aqueous polymer systems, Biochim. Biophys. Acta 640:557 (1981)PubMedCrossRefGoogle Scholar
  10. 10.
    D.J.L. McIver and S. Schürch, Interfacial free energies of intact and reconstituted erythrocyte surfaces; implications for biological adhesion, Biochim. Biophys. Acta 691:52 (1983)Google Scholar
  11. 11.
    S.A. Johnstone, S. Schürch, D.J.L. McIver, E.A. Jacobson and E.R. Tustanoff, Membrane glycoprotein and surface free energy changes in hypoxic fibroblast cells, Biochim. Biophys. Acta 815:159 (1985)PubMedCrossRefGoogle Scholar
  12. 12.
    D.J.L. McIver and S. Schürch, Free energies at the biosurface-water interface, in: “The Biophysics of Water,” F. Franks and S. Mathias, eds., Jonathan Wiley, London (1983)Google Scholar
  13. 13.
    A. Boyum, Isolation of lymphocytes, granulocytes and macrophages, Scand. J. Immunol. 5, Supp. 5:9 (1976)PubMedCrossRefGoogle Scholar
  14. 14.
    Y. Rotenburg, L. Boruvka and A.W. Neumann, Determination of surface tension and contact angle from the shapes of axisymmetric fluid interfaces, J. Colloid Int. Sci. 93:169 (1983)CrossRefGoogle Scholar
  15. 15.
    D.J.L. McIver, K.A. Sharp, D.E. Brooks and E.A. Evans, unpublished resultsGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Donald J. L. McIver
    • 1
  • Samuel Schürch
    • 2
  1. 1.Departments of Pharmacology and Toxicology, Medical Biophysics, Medicine and The Robarts Research InstituteUniversity of Western OntarioLondonCanada
  2. 2.Faculty of MedicineUniversity of CalgaryCalgaryCanada

Personalised recommendations