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. 1986 Dec;50(6):1117–1126. doi: 10.1016/S0006-3495(86)83556-1

Interaction forces between red cells agglutinated by antibody. II. Measurement of hydrodynamic force of breakup.

S P Tha, J Shuster, H L Goldsmith
PMCID: PMC1329786  PMID: 3801572

Abstract

The expressions derived in the previous paper for the respective normal, F3, and shear forces, Fshear, acting along and perpendicular to the axis of a doublet of rigid spheres, were used to determine the hydrodynamic forces required to separate two red cell spheres of antigenic type B crosslinked by the corresponding antibody. Cells were sphered and swollen in isotonic buffered glycerol containing 8 X 10(-5) M sodium dodecyl sulfate, fixed in 0.085% glutaraldehyde, and suspended in aqueous glycerol (viscosity: 15-34 mPa s), containing 0.15 M NaCl and anti-B antibody from human hyperimmune antiserum at concentrations from 0.73 to 3.56 vol%. After incubating and mixing for 12 h, doublets were observed through a microscope flowing in a 178-micron tube by gravity feed between two reservoirs. Using a traveling microtube apparatus, the doublets were tracked in a constantly accelerating flow and the translational and rotational motions were recorded on videotape until breakup occurred. From a frame by frame replay of the tape, the radial position, velocity and orientation of the doublet were obtained and the normal and shear forces of separation at breakup computed. Both forces increased significantly with increasing antiserum concentration, the mean values of F3 increasing from 0.060 to 0.197 nN, and Fshear from 0.023 to 0.072 nN. There was no significant effect of glycerol viscosity on the forces of separation. It was not possible to determine whether the shear or normal force was responsible for doublet separation. Measurements of the mean dimensionless period of rotation, TG, of doublets in suspensions containing 0.73 and 2.40% antiserum undergoing steady flow were also made to test whether the spheres were rigidly linked or capable of some independent rotation. A fairly narrow distribution in TG about the value 15.64, predicted for rigidly-linked doublets, was obtained at both antiserum concentrations.

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  1. Bell G. I. Models for the specific adhesion of cells to cells. Science. 1978 May 12;200(4342):618–627. doi: 10.1126/science.347575. [DOI] [PubMed] [Google Scholar]
  2. Capo C., Garrouste F., Benoliel A. M., Bongrand P., Ryter A., Bell G. I. Concanavalin-A-mediated thymocyte agglutination: a model for a quantitative study of cell adhesion. J Cell Sci. 1982 Aug;56:21–48. doi: 10.1242/jcs.56.1.21. [DOI] [PubMed] [Google Scholar]
  3. Economidou J., Hughes-Jones N. C., Gardner B. Quantitative measurements concerning A and B antigen sites. Vox Sang. 1967 May;12(5):321–328. doi: 10.1111/j.1423-0410.1967.tb03362.x. [DOI] [PubMed] [Google Scholar]
  4. Evans E. A. Detailed mechanics of membrane-membrane adhesion and separation. I. Continuum of molecular cross-bridges. Biophys J. 1985 Jul;48(1):175–183. doi: 10.1016/S0006-3495(85)83770-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans E. A. Detailed mechanics of membrane-membrane adhesion and separation. II. Discrete kinetically trapped molecular cross-bridges. Biophys J. 1985 Jul;48(1):185–192. doi: 10.1016/S0006-3495(85)83771-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans E. A., Parsegian V. A. Energetics of membrane deformation and adhesion in cell and vesicle aggregation. Ann N Y Acad Sci. 1983;416:13–33. doi: 10.1111/j.1749-6632.1983.tb35176.x. [DOI] [PubMed] [Google Scholar]
  7. Evans E., Leung A. Adhesivity and rigidity of erythrocyte membrane in relation to wheat germ agglutinin binding. J Cell Biol. 1984 Apr;98(4):1201–1208. doi: 10.1083/jcb.98.4.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goldsmith H. L., Gold P., Shuster J., Takamura K. Interactions between sphered human red cells in tube flow: technique for measuring the strength of antigen--antibody bonds. Microvasc Res. 1982 Mar;23(2):231–238. doi: 10.1016/0026-2862(82)90067-x. [DOI] [PubMed] [Google Scholar]
  9. Goldsmith H. L., Lichtarge O., Tessier-Lavigne M., Spain S. Some model experiments in hemodynamics: VI. Two-body collisions between blood cells. Biorheology. 1981;18(3-6):531–555. doi: 10.3233/bir-1981-183-617. [DOI] [PubMed] [Google Scholar]
  10. Goldsmith H. L., Mason S. G. Some model experiments in hemodynamics-V: microrheological techniques. Biorheology. 1975 Jun;12(3-4):181–192. doi: 10.3233/bir-1975-123-408. [DOI] [PubMed] [Google Scholar]
  11. Hakomori S. Blood group ABH and Ii antigens of human erythrocytes: chemistry, polymorphism, and their developmental change. Semin Hematol. 1981 Jan;18(1):39–62. [PubMed] [Google Scholar]
  12. Hughes-Jones N. C. The attachment of IgG molecules on the red cell surface. Haematologia (Budap) 1972;6(3):269–274. [PubMed] [Google Scholar]
  13. Knox R. J., Nordt F. J., Seaman G. V., Brooks D. E. Rheology of erythrocyte suspensions: dextran-mediated aggregation of deformable and nondeformable erythrocytes. Biorheology. 1977;14(2-3):75–84. doi: 10.3233/bir-1977-142-303. [DOI] [PubMed] [Google Scholar]
  14. Lenard J., Singer S. J. Alteration of the conformation of proteins in red blood cell membranes and in solution by fixatives used in electron microscopy. J Cell Biol. 1968 Apr;37(1):117–121. doi: 10.1083/jcb.37.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marquardt M. D., Gordon J. A. Glutaraldehyde fixation and the mechanism of erythroycte agglutination by concanavalin A and soybean agglutinin. Exp Cell Res. 1975 Mar 15;91(2):310–316. doi: 10.1016/0014-4827(75)90109-3. [DOI] [PubMed] [Google Scholar]
  16. Peters K., Richards F. M. Chemical cross-linking: reagents and problems in studies of membrane structure. Annu Rev Biochem. 1977;46:523–551. doi: 10.1146/annurev.bi.46.070177.002515. [DOI] [PubMed] [Google Scholar]
  17. Sine S. M., Steinbach J. H. Activation of a nicotinic acetylcholine receptor. Biophys J. 1984 Jan;45(1):175–185. doi: 10.1016/S0006-3495(84)84146-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Trommler A., Gingell D., Wolf H. Red blood cells experience electrostatic repulsion but make molecular adhesions with glass. Biophys J. 1985 Nov;48(5):835–841. doi: 10.1016/S0006-3495(85)83842-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vassar P. S., Hards J. M., Brooks D. E., Hagenberger B., Seaman G. V. Physicochemical effects of aldehydes on the human erythrocyte. J Cell Biol. 1972 Jun;53(3):809–818. doi: 10.1083/jcb.53.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. van Oss C. J., Absolom D. R. Zeta potentials, van der Waals forces and hemagglutination. Vox Sang. 1983 Mar;44(3):183–190. doi: 10.1111/j.1423-0410.1983.tb01883.x. [DOI] [PubMed] [Google Scholar]

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