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. 1985 Mar 1;100(3):860–872. doi: 10.1083/jcb.100.3.860

Lymphocyte mechanical response triggered by cross-linking surface receptors

PMCID: PMC2113522  PMID: 4038710

Abstract

Using a recently developed method (Petersen, N. O., W. B. McConnaughey, and E. L. Elson, 1982, Proc. Natl. Acad. Sci. USA., 79:5327-5331), we have measured changes in the deformability of lymphocytes triggered by cross-linking cell surface proteins. Our study was motivated by two previously demonstrated phenomena: the redistribution ("capping") of cross-linked surface immunoglobulin (sIg) on B lymphocytes and the inhibition of capping and lateral diffusion ("anchorage modulation") of sIg by the tetravalent lectin Concanavalin A (Con A). Both capping and anchorage modulation are initiated by cross-linking cell surface proteins and both require participation of the cytoskeleton. We have shown that the resistance of lymphocytes to deformation strongly increased when sIg or Con A acceptors were cross-linked. We have measured changes in deformability in terms of an empirical "stiffness" parameter, defined as the rate at which the force of cellular compression increases with the extent of compression. For untreated cells the stiffness was approximately 0.15 mdyn/micron; for cells treated with antibodies against sIg or with Con A the stiffness increased to approximately 0.6 or 0.4 mdyn/micron, respectively. The stiffness decreased after completion of the capping of sIg. The increases in stiffness could be reversed to various extents by cytochalasin D and by colchicine. The need for cross-linking was demonstrated by the failure both of monovalent Fab' fragments of the antibodies against sIg and of succinylated Con A (a poor cross-linker) to cause an increase in stiffness. We conclude that capping and anchorage modulation involve changes in the lymphocyte cytoskeleton and possibly other cytoplasmic properties, which increase the cellular viscoelastic resistance to deformation. Similar increases in cell stiffness could be produced by exposing cells to hypertonic medium, azide ions, and to a calcium ionophore in the presence of calcium ions. These results shed new light on the capabilities of the lymphocyte cytoskeleton and its role in capping and anchorage modulation. They also demonstrate that measurements of cellular deformability can characterize changes in cytoskeletal functions initiated by signals originating at the cell surface.

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Selected References

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  1. Bershadsky A. D., Gelfand V. I., Svitkina T. M., Tint I. S. Destruction of microfilament bundles in mouse embryo fibroblasts treated with inhibitors of energy metabolism. Exp Cell Res. 1980 Jun;127(2):421–429. doi: 10.1016/0014-4827(80)90446-2. [DOI] [PubMed] [Google Scholar]
  2. Bourguignon L. Y., Nagpal M. L., Hsing Y. C. Phosphorylation of myosin light chain during capping of mouse T-lymphoma cells. J Cell Biol. 1981 Dec;91(3 Pt 1):889–894. doi: 10.1083/jcb.91.3.889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bourguignon L. Y., Singer S. J. Transmembrane interactions and the mechanism of capping of surface receptors by their specific ligands. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5031–5035. doi: 10.1073/pnas.74.11.5031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braun J., Fujiwara K., Pollard T. D., Unanue E. R. Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. I. Relationship to cytoplasmic myosin. J Cell Biol. 1978 Nov;79(2 Pt 1):409–418. doi: 10.1083/jcb.79.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braun J., Fujiwara K., Pollard T. D., Unanue E. R. Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. II. Contrasting effects of local anesthetics and a calcium ionophore. J Cell Biol. 1978 Nov;79(2 Pt 1):419–426. doi: 10.1083/jcb.79.2.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Braun J., Hochman P. S., Unanue E. R. Ligand-induced association of surface immunoglobulin with the detergent-insoluble cytoskeletal matrix of the B lymphocyte. J Immunol. 1982 Mar;128(3):1198–1204. [PubMed] [Google Scholar]
  7. Braun J., Sha'afi R. I., Unanue E. R. Crosslinking by ligands to surface immunoglobulin triggers mobilization of intracellular 45Ca2+ in B lymphocytes. J Cell Biol. 1979 Sep;82(3):755–766. doi: 10.1083/jcb.82.3.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bretscher M. S. Endocytosis: relation to capping and cell locomotion. Science. 1984 May 18;224(4650):681–686. doi: 10.1126/science.6719108. [DOI] [PubMed] [Google Scholar]
  9. Daily B., Elson E. L., Zahalak G. I. Cell poking. Determination of the elastic area compressibility modulus of the erythrocyte membrane. Biophys J. 1984 Apr;45(4):671–682. doi: 10.1016/S0006-3495(84)84209-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dragsten P., Henkart P., Blumenthal R., Weinstein J., Schlessinger J. Lateral diffusion of surface immunoglobulin, Thy-1 antigen, and a lipid probe in lymphocyte plasma membranes. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5163–5167. doi: 10.1073/pnas.76.10.5163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Fechheimer M., Cebra J. J. Phosphorylation of lymphocyte myosin catalyzed in vitro and in intact cells. J Cell Biol. 1982 May;93(2):261–268. doi: 10.1083/jcb.93.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flanagan J., Koch G. L. Cross-linked surface Ig attaches to actin. Nature. 1978 May 25;273(5660):278–281. doi: 10.1038/273278a0. [DOI] [PubMed] [Google Scholar]
  15. Gabbiani G., Chaponnier C., Zumbe A., Vassalli P. Actin and tubulin co-cap with surface immunoglobulins in mouse B lymphocytes. Nature. 1977 Oct 20;269(5630):697–698. doi: 10.1038/269697a0. [DOI] [PubMed] [Google Scholar]
  16. Geiger B., Singer S. J. The participation of alpha-actinin in the capping of cell membrane components. Cell. 1979 Jan;16(1):213–222. doi: 10.1016/0092-8674(79)90202-2. [DOI] [PubMed] [Google Scholar]
  17. Henis Y. I., Elson E. L. Differences in the response of several cell types to inhibition of surface receptor mobility by local concanavalin A binding. Exp Cell Res. 1981 Nov;136(1):189–201. doi: 10.1016/0014-4827(81)90050-1. [DOI] [PubMed] [Google Scholar]
  18. Henis Y. I., Elson E. L. Inhibition of the mobility of mouse lymphocyte surface immunoglobulins by locally bound concanavalin A. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1072–1076. doi: 10.1073/pnas.78.2.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoessli D., Rungger-Brändle E., Jockusch B. M., Gabbiani G. Lymphocyte alpha-actinin. Relationship to cell membrane and co-capping with surface receptors. J Cell Biol. 1980 Feb;84(2):305–314. doi: 10.1083/jcb.84.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kerrick W. G., Bourguignon L. Y. Regulation of receptor capping in mouse lymphoma T cells by Ca2+-activated myosin light chain kinase. Proc Natl Acad Sci U S A. 1984 Jan;81(1):165–169. doi: 10.1073/pnas.81.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Korn E. D. Actin polymerization and its regulation by proteins from nonmuscle cells. Physiol Rev. 1982 Apr;62(2):672–737. doi: 10.1152/physrev.1982.62.2.672. [DOI] [PubMed] [Google Scholar]
  22. Laub F., Kaplan M., Gitler C. Actin polymerization accompanies Thy-1-capping on mouse thymocytes. FEBS Lett. 1981 Feb 9;124(1):35–38. doi: 10.1016/0014-5793(81)80048-8. [DOI] [PubMed] [Google Scholar]
  23. Levine J., Willard M. Redistribution of fodrin (a component of the cortical cytoplasm) accompanying capping of cell surface molecules. Proc Natl Acad Sci U S A. 1983 Jan;80(1):191–195. doi: 10.1073/pnas.80.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nelson G. A., Andrews M. L., Karnovsky M. J. Participation of calmodulin in immunoglobulin capping. J Cell Biol. 1982 Dec;95(3):771–780. doi: 10.1083/jcb.95.3.771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nelson W. J., Colaço C. A., Lazarides E. Involvement of spectrin in cell-surface receptor capping in lymphocytes. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1626–1630. doi: 10.1073/pnas.80.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Norberg B. The formation of an ATP-induced constriction ring in a glycerinated lymphocyte during migration. Scand J Haematol. 1971;8(1):75–80. doi: 10.1111/j.1600-0609.1971.tb02675.x. [DOI] [PubMed] [Google Scholar]
  27. Oliver J. M., Berlin R. D. Mechanisms that regulate the structural and functional architecture of cell surfaces. Int Rev Cytol. 1982;74:55–94. doi: 10.1016/s0074-7696(08)61169-9. [DOI] [PubMed] [Google Scholar]
  28. Parham P., Androlewicz M. J., Brodsky F. M., Holmes N. J., Ways J. P. Monoclonal antibodies: purification, fragmentation and application to structural and functional studies of class I MHC antigens. J Immunol Methods. 1982 Sep 17;53(2):133–173. doi: 10.1016/0022-1759(82)90137-5. [DOI] [PubMed] [Google Scholar]
  29. Petersen N. O., McConnaughey W. B., Elson E. L. Dependence of locally measured cellular deformability on position on the cell, temperature, and cytochalasin B. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5327–5331. doi: 10.1073/pnas.79.17.5327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pozzan T., Arslan P., Tsien R. Y., Rink T. J. Anti-immunoglobulin, cytoplasmic free calcium, and capping in B lymphocytes. J Cell Biol. 1982 Aug;94(2):335–340. doi: 10.1083/jcb.94.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pozzan T., Corps A. N., Montecucco C., Hesketh T. R., Metcalfe J. C. Cap formation by various ligands on lymphocytes shows the same dependence on high cellular ATP levels. Biochim Biophys Acta. 1980 Nov 18;602(3):558–566. doi: 10.1016/0005-2736(80)90334-x. [DOI] [PubMed] [Google Scholar]
  32. Rink T. J., Montecucco C., Hesketh T. R., Tsien R. Y. Lymphocyte membrane potential assessed with fluorescent probes. Biochim Biophys Acta. 1980;595(1):15–30. doi: 10.1016/0005-2736(80)90243-6. [DOI] [PubMed] [Google Scholar]
  33. Rutishauser U., Yahara I., Edelman G. M. Morphology, motility, and surface behavior of lymphocytes bound to nylon fibers. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1149–1153. doi: 10.1073/pnas.71.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ryan J. L., Arbeit R. D., Dickler H. B., Henkart P. A. Inhibition of lymphocyte mitogenesis by immobilized antigen-antibody complexes. J Exp Med. 1975 Oct 1;142(4):814–826. doi: 10.1084/jem.142.4.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Salisbury J. L., Condeelis J. S., Maihle N. J., Satir P. Calmodulin localization during capping and receptor-mediated endocytosis. Nature. 1981 Nov 12;294(5837):163–166. doi: 10.1038/294163a0. [DOI] [PubMed] [Google Scholar]
  36. Schmid-Schönbein G. W., Shih Y. Y., Chien S. Morphometry of human leukocytes. Blood. 1980 Nov;56(5):866–875. [PubMed] [Google Scholar]
  37. Schreiner G. F., Fujiwara K., Pollard T. D., Unanue E. R. Redistribution of myosin accompanying capping of surface Ig. J Exp Med. 1977 May 1;145(5):1393–1398. doi: 10.1084/jem.145.5.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schreiner G. F., Unanue E. R. Calcium-sensitive modulation of Ig capping: evidence supporting a cytoplasmic control of ligand-receptor complexes. J Exp Med. 1976 Jan 1;143(1):15–31. doi: 10.1084/jem.143.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schreiner G. F., Unanue E. R. Membrane and cytoplasmic changes in B lymphocytes induced by ligand-surface immunoglobulin interaction. Adv Immunol. 1976;24:37–165. doi: 10.1016/s0065-2776(08)60329-6. [DOI] [PubMed] [Google Scholar]
  40. Weeds A. Actin-binding proteins--regulators of cell architecture and motility. Nature. 1982 Apr 29;296(5860):811–816. doi: 10.1038/296811a0. [DOI] [PubMed] [Google Scholar]
  41. Yahara I., Edelman G. M. Modulation of lymphocyte receptor mobility by locally bound concanavalin A. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1579–1583. doi: 10.1073/pnas.72.4.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yahara I., Edelman G. M. Modulation of lymphocyte receptor redistribution by concanavalin A, anti-mitotic agents and alterations of pH. Nature. 1973 Nov 16;246(5429):152–155. doi: 10.1038/246152a0. [DOI] [PubMed] [Google Scholar]
  43. Yahara I., Edelman G. M. Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A. Proc Natl Acad Sci U S A. 1972 Mar;69(3):608–612. doi: 10.1073/pnas.69.3.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yahara I., Edelman G. M. The effects of concanavalin A on the mobility of lymphocyte surface receptors. Exp Cell Res. 1973 Sep;81(1):143–155. doi: 10.1016/0014-4827(73)90121-3. [DOI] [PubMed] [Google Scholar]
  45. Yahara I., Kakimoto-Sameshima F. Analysis of ligand-independent cap formation induced in hypertonic medium. Exp Cell Res. 1979 Mar 15;119(2):237–252. doi: 10.1016/0014-4827(79)90352-5. [DOI] [PubMed] [Google Scholar]
  46. Yahara I., Kakimoto-Sameshima F. Ligand-independent cap formation: redistribution of surface receptors on mouse lymphocytes and thymocytes in hypertonic medium. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4511–4515. doi: 10.1073/pnas.74.10.4511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. de Petris S. Concanavalin A receptors, immunoglobulins, and theta antigen of the lymphocyte surface. Interactions with concanavalin A and with Cytoplasmic structures. J Cell Biol. 1975 Apr;65(1):123–146. doi: 10.1083/jcb.65.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]

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