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. 1992 Feb 2;116(4):997–1006. doi: 10.1083/jcb.116.4.997

Micromanipulation of adhesion of a Jurkat cell to a planar bilayer membrane containing lymphocyte function-associated antigen 3 molecules

PMCID: PMC2289335  PMID: 1370839

Abstract

Cell adhesion plays a fundamental role in the organization of cells in differentiated organs, cell motility, and immune response. A novel micromanipulation method is employed to quantify the direct contribution of surface adhesion receptors to the physical strength of cell adhesion. In this technique, a cell is brought into contact with a glass-supported planar membrane reconstituted with a known concentration of a given type of adhesion molecules. After a period of incubation (5-10 min), the cell is detached from the planar bilayer by pulling away the pipette holding the cell in the direction perpendicular to the glass-supported planar bilayer. In particular, we investigated the adhesion between a Jurkat cell expressing CD2 and a glass-supported planar bilayer containing either the glycosyl- phosphatidylinositol (GPI) or the transmembrane (TM) isoform of the counter-receptor lymphocyte function-associated antigen 3 (LFA-3) at a concentration of 1,000 molecules/microns 2. In response to the pipette force the Jurkat cells that adhered to the planar bilayer containing the GPI isoform of LFA-3 underwent extensive elongation. When the contact radius was reduced by approximately 50%, the cell then detached quickly from its substrate. The aspiration pressure required to detach a Jurkat cell from its substrate was comparable to that required to detach a cytotoxic T cell from its target cell. Jurkat cells that had been separated from the substrate again adhered strongly to the planar bilayer when brought to proximity by micromanipulation. In experiments using the planar bilayer containing the TM isoform of LFA-3, Jurkat cells detached with little resistance to micromanipulation and without changing their round shape.

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Selected References

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  1. Bell G. I., Dembo M., Bongrand P. Cell adhesion. Competition between nonspecific repulsion and specific bonding. Biophys J. 1984 Jun;45(6):1051–1064. doi: 10.1016/S0006-3495(84)84252-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Brian A. A., McConnell H. M. Allogeneic stimulation of cytotoxic T cells by supported planar membranes. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6159–6163. doi: 10.1073/pnas.81.19.6159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carpén O., Dustin M. L., Springer T. A., Swafford J. A., Beckett L. A., Caulfield J. P. Motility and ultrastructure of large granular lymphocytes on lipid bilayers reconstituted with adhesion receptors LFA-1, ICAM-1, and two isoforms of LFA-3. J Cell Biol. 1991 Nov;115(3):861–871. doi: 10.1083/jcb.115.3.861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chan P. Y., Lawrence M. B., Dustin M. L., Ferguson L. M., Golan D. E., Springer T. A. Influence of receptor lateral mobility on adhesion strengthening between membranes containing LFA-3 and CD2. J Cell Biol. 1991 Oct;115(1):245–255. doi: 10.1083/jcb.115.1.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chao N. M., Young S. H., Poo M. M. Localization of cell membrane components by surface diffusion into a "trap". Biophys J. 1981 Oct;36(1):139–153. doi: 10.1016/S0006-3495(81)84721-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dembo M., Torney D. C., Saxman K., Hammer D. The reaction-limited kinetics of membrane-to-surface adhesion and detachment. Proc R Soc Lond B Biol Sci. 1988 Jun 22;234(1274):55–83. doi: 10.1098/rspb.1988.0038. [DOI] [PubMed] [Google Scholar]
  8. Dong C., Skalak R., Sung K. L., Schmid-Schönbein G. W., Chien S. Passive deformation analysis of human leukocytes. J Biomech Eng. 1988 Feb;110(1):27–36. doi: 10.1115/1.3108402. [DOI] [PubMed] [Google Scholar]
  9. Duband J. L., Nuckolls G. H., Ishihara A., Hasegawa T., Yamada K. M., Thiery J. P., Jacobson K. Fibronectin receptor exhibits high lateral mobility in embryonic locomoting cells but is immobile in focal contacts and fibrillar streaks in stationary cells. J Cell Biol. 1988 Oct;107(4):1385–1396. doi: 10.1083/jcb.107.4.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dustin M. L., Olive D., Springer T. A. Correlation of CD2 binding and functional properties of multimeric and monomeric lymphocyte function-associated antigen 3. J Exp Med. 1989 Feb 1;169(2):503–517. doi: 10.1084/jem.169.2.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dustin M. L., Sanders M. E., Shaw S., Springer T. A. Purified lymphocyte function-associated antigen 3 binds to CD2 and mediates T lymphocyte adhesion. J Exp Med. 1987 Mar 1;165(3):677–692. doi: 10.1084/jem.165.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dustin M. L., Selvaraj P., Mattaliano R. J., Springer T. A. Anchoring mechanisms for LFA-3 cell adhesion glycoprotein at membrane surface. 1987 Oct 29-Nov 4Nature. 329(6142):846–848. doi: 10.1038/329846a0. [DOI] [PubMed] [Google Scholar]
  13. Dustin M. L., Springer T. A. T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1. Nature. 1989 Oct 19;341(6243):619–624. doi: 10.1038/341619a0. [DOI] [PubMed] [Google Scholar]
  14. Evans E., Buxbaum K. Affinity of red blood cell membrane for particle surfaces measured by the extent of particle encapsulation. Biophys J. 1981 Apr;34(1):1–12. doi: 10.1016/S0006-3495(81)84834-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Evans E., Yeung A. Apparent viscosity and cortical tension of blood granulocytes determined by micropipet aspiration. Biophys J. 1989 Jul;56(1):151–160. doi: 10.1016/S0006-3495(89)82660-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gallik S., Usami S., Jan K. M., Chien S. Shear stress-induced detachment of human polymorphonuclear leukocytes from endothelial cell monolayers. Biorheology. 1989;26(4):823–834. doi: 10.3233/bir-1989-26413. [DOI] [PubMed] [Google Scholar]
  18. Hollander N., Selvaraj P., Springer T. A. Biosynthesis and function of LFA-3 in human mutant cells deficient in phosphatidylinositol-anchored proteins. J Immunol. 1988 Dec 15;141(12):4283–4290. [PubMed] [Google Scholar]
  19. Kupfer A., Singer S. J. Cell biology of cytotoxic and helper T cell functions: immunofluorescence microscopic studies of single cells and cell couples. Annu Rev Immunol. 1989;7:309–337. doi: 10.1146/annurev.iy.07.040189.001521. [DOI] [PubMed] [Google Scholar]
  20. Lawrence M. B., Springer T. A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991 May 31;65(5):859–873. doi: 10.1016/0092-8674(91)90393-d. [DOI] [PubMed] [Google Scholar]
  21. McClay D. R., Ettensohn C. A. Cell adhesion in morphogenesis. Annu Rev Cell Biol. 1987;3:319–345. doi: 10.1146/annurev.cb.03.110187.001535. [DOI] [PubMed] [Google Scholar]
  22. McCloskey M. A., Poo M. M. Contact-induced redistribution of specific membrane components: local accumulation and development of adhesion. J Cell Biol. 1986 Jun;102(6):2185–2196. doi: 10.1083/jcb.102.6.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McConnell H. M., Watts T. H., Weis R. M., Brian A. A. Supported planar membranes in studies of cell-cell recognition in the immune system. Biochim Biophys Acta. 1986 Jun 12;864(1):95–106. doi: 10.1016/0304-4157(86)90016-x. [DOI] [PubMed] [Google Scholar]
  24. Mohandas N., Hochmuth R. M., Spaeth E. E. Adhesion of red cells to foreign surfaces in the presence of flow. J Biomed Mater Res. 1974 Mar;8(2):119–136. doi: 10.1002/jbm.820080203. [DOI] [PubMed] [Google Scholar]
  25. Plunkett M. L., Springer T. A. Purification and characterization of the lymphocyte function-associated-2 (LFA-2) molecule. J Immunol. 1986 Jun 1;136(11):4181–4187. [PubMed] [Google Scholar]
  26. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  27. Selvaraj P., Dustin M. L., Mitnacht R., Hünig T., Springer T. A., Plunkett M. L. Rosetting of human T lymphocytes with sheep and human erythrocytes. Comparison of human and sheep ligand binding using purified E receptor. J Immunol. 1987 Oct 15;139(8):2690–2695. [PubMed] [Google Scholar]
  28. Shaw S., Luce G. E., Quinones R., Gress R. E., Springer T. A., Sanders M. E. Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones. Nature. 1986 Sep 18;323(6085):262–264. doi: 10.1038/323262a0. [DOI] [PubMed] [Google Scholar]
  29. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  30. Springer T. A. Adhesion receptors of the immune system. Nature. 1990 Aug 2;346(6283):425–434. doi: 10.1038/346425a0. [DOI] [PubMed] [Google Scholar]
  31. Sung K. L., Dong C., Schmid-Schönbein G. W., Chien S., Skalak R. Leukocyte relaxation properties. Biophys J. 1988 Aug;54(2):331–336. doi: 10.1016/S0006-3495(88)82963-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sung K. L., Sung L. A., Crimmins M., Burakoff S. J., Chien S. Determination of junction avidity of cytolytic T cell and target cell. Science. 1986 Dec 12;234(4782):1405–1408. doi: 10.1126/science.3491426. [DOI] [PubMed] [Google Scholar]
  33. Sung K. L., Sung L. A., Crimmins M., Burakoff S. J., Chien S. Dynamic changes in viscoelastic properties in cytotoxic T-lymphocyte-mediated killing. J Cell Sci. 1988 Oct;91(Pt 2):179–189. doi: 10.1242/jcs.91.2.179. [DOI] [PubMed] [Google Scholar]
  34. Tamm L. K. Lateral diffusion and fluorescence microscope studies on a monoclonal antibody specifically bound to supported phospholipid bilayers. Biochemistry. 1988 Mar 8;27(5):1450–1457. doi: 10.1021/bi00405a009. [DOI] [PubMed] [Google Scholar]
  35. Tozeren A., Sung K. L., Chien S. Theoretical and experimental studies on cross-bridge migration during cell disaggregation. Biophys J. 1989 Mar;55(3):479–487. doi: 10.1016/S0006-3495(89)82841-3. [DOI] [PMC free article] [PubMed] [Google Scholar]

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