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. 1987 Mar 1;165(3):677–692. doi: 10.1084/jem.165.3.677

Purified lymphocyte function-associated antigen 3 binds to CD2 and mediates T lymphocyte adhesion

PMCID: PMC2188278  PMID: 3102676

Abstract

CD2 is a T lymphocyte glycoprotein that functions in adhesion of T lymphocytes and also as a putative receptor for activation signals. Functional data suggest that LFA-3, a widely distributed cell surface glycoprotein, may be the biological ligand of CD2. We have purified LFA- 3 from human erythrocytes and characterized the purified protein functionally. LFA-3 bound specifically to CD2+ cells, and this binding was inhibited by CD2 mAb. Conversely, purified LFA-3 inhibited binding of CD2 mAb to cells, and the concentration required for this effect suggests that LFA-3 half-saturated CD2 at 1-5 nM LFA-3. Purified LFA-3 inhibited rosetting of human and sheep erythrocytes with CD2+ T lymphoma cells and T lymphocytes, and mediated aggregation of a CD2+ T lymphoma cell line. Purified LFA-3 reconstituted into planar membranes mediated efficient CD2-dependent adhesion of T lymphoblasts. These data demonstrate that LFA-3 is a ligand for CD2 and that LFA-3 can mediate T lymphocyte adhesion.

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Selected References

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  1. Baxley G., Bishop G. B., Cooper A. G., Wortis H. H. Rosetting of human red blood cells to thymocytes and thymus-derived cells. Clin Exp Immunol. 1973 Nov;15(3):385–392. [PMC free article] [PubMed] [Google Scholar]
  2. Bentwich Z., Kunkel H. G. Specific properties of human B and T lymphocytes and alterations in disease. Transplant Rev. 1973;16:29–50. doi: 10.1111/j.1600-065x.1973.tb00116.x. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Despont J. P., Abel C. A., Grey H. M. Sialic acids and sialyltransferases in murine lymphoid cells: indicators of T cell maturation. Cell Immunol. 1975 Jun;17(2):487–494. doi: 10.1016/s0008-8749(75)80052-9. [DOI] [PubMed] [Google Scholar]
  6. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  7. Gay D., Coeshott C., Golde W., Kappler J., Marrack P. The major histocompatibility complex-restricted antigen receptor on T cells. IX. Role of accessory molecules in recognition of antigen plus isolated IA. J Immunol. 1986 Mar 15;136(6):2026–2032. [PubMed] [Google Scholar]
  8. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  9. Haynes B. F. The role of the thymic microenvironment in promotion of early stages of human T cell maturation. Clin Res. 1986 Sep;34(3):422–431. [PubMed] [Google Scholar]
  10. Holter W., Fischer G. F., Majdic O., Stockinger H., Knapp W. T cell stimulation via the erythrocyte receptor. Synergism between monoclonal antibodies and phorbol myristate acetate without changes of free cytoplasmic Ca++ levels. J Exp Med. 1986 Mar 1;163(3):654–664. doi: 10.1084/jem.163.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hünig T. R. The ligand of the erythrocyte receptor of T lymphocytes: expression on white blood cells and possible involvement in T cell activation. J Immunol. 1986 Mar 15;136(6):2103–2108. [PubMed] [Google Scholar]
  12. Hünig T. The cell surface molecule recognized by the erythrocyte receptor of T lymphocytes. Identification and partial characterization using a monoclonal antibody. J Exp Med. 1985 Sep 1;162(3):890–901. doi: 10.1084/jem.162.3.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Janiak M. J., Small D. M., Shipley G. G. Temperature and compositional dependence of the structure of hydrated dimyristoyl lecithin. J Biol Chem. 1979 Jul 10;254(13):6068–6078. [PubMed] [Google Scholar]
  14. Krensky A. M., Robbins E., Springer T. A., Burakoff S. J. LFA-1, LFA-2, and LFA-3 antigens are involved in CTL-target conjugation. J Immunol. 1984 May;132(5):2180–2182. [PubMed] [Google Scholar]
  15. Krensky A. M., Sanchez-Madrid F., Robbins E., Nagy J. A., Springer T. A., Burakoff S. J. The functional significance, distribution, and structure of LFA-1, LFA-2, and LFA-3: cell surface antigens associated with CTL-target interactions. J Immunol. 1983 Aug;131(2):611–616. [PubMed] [Google Scholar]
  16. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  17. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  18. Martin P. J., Longton G., Ledbetter J. A., Newman W., Braun M. P., Beatty P. G., Hansen J. A. Identification and functional characterization of two distinct epitopes on the human T cell surface protein Tp50. J Immunol. 1983 Jul;131(1):180–185. [PubMed] [Google Scholar]
  19. Meuer S. C., Hussey R. E., Fabbi M., Fox D., Acuto O., Fitzgerald K. A., Hodgdon J. C., Protentis J. P., Schlossman S. F., Reinherz E. L. An alternative pathway of T-cell activation: a functional role for the 50 kd T11 sheep erythrocyte receptor protein. Cell. 1984 Apr;36(4):897–906. doi: 10.1016/0092-8674(84)90039-4. [DOI] [PubMed] [Google Scholar]
  20. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  21. Nalet V., Fournier C. Human autologous rosette-forming cells. III. Binding of erythrocytes from different species to the T-cell receptors for autologous red blood cells. Cell Immunol. 1985 Nov;96(1):126–136. doi: 10.1016/0008-8749(85)90345-4. [DOI] [PubMed] [Google Scholar]
  22. Palacios R., Llorente L., Alarcón-Segovia D., Ruíz-Arguelles A., Díaz-Jouanen E. Autologous rosette-forming T cells as the responding cells in human autologous mixed-lymphocyte reaction. J Clin Invest. 1980 Jun;65(6):1527–1530. doi: 10.1172/JCI109819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Palacios R., Martinez-Maza O. Is the E receptor on human T lymphocytes a "negative signal receptor"? J Immunol. 1982 Dec;129(6):2479–2485. [PubMed] [Google Scholar]
  24. Plunkett M. L., Sanders M. E., Selvaraj P., Dustin M. L., Springer T. A. Rosetting of activated human T lymphocytes with autologous erythrocytes. Definition of the receptor and ligand molecules as CD2 and lymphocyte function-associated antigen 3 (LFA-3). J Exp Med. 1987 Mar 1;165(3):664–676. doi: 10.1084/jem.165.3.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reed J. C., Tadmori W., Kamoun M., Koretzky G., Nowell P. C. Suppression of interleukin 2 receptor acquisition by monoclonal antibodies recognizing the 50 KD protein associated with the sheep erythrocyte receptor on human T lymphocytes. J Immunol. 1985 Mar;134(3):1631–1639. [PubMed] [Google Scholar]
  26. Rothlein R., Dustin M. L., Marlin S. D., Springer T. A. A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1. J Immunol. 1986 Aug 15;137(4):1270–1274. [PubMed] [Google Scholar]
  27. Sanchez-Madrid F., Krensky A. M., Ware C. F., Robbins E., Strominger J. L., Burakoff S. J., Springer T. A. Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7489–7493. doi: 10.1073/pnas.79.23.7489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanchez-Madrid F., Nagy J. A., Robbins E., Simon P., Springer T. A. A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1), and the p150,95 molecule. J Exp Med. 1983 Dec 1;158(6):1785–1803. doi: 10.1084/jem.158.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Siliciano R. F., Hemesath T. J., Pratt J. C., Dintzis R. Z., Dintzis H. M., Acuto O., Shin H. S., Reinherz E. L. Direct evidence for the existence of nominal antigen binding sites on T cell surface Ti alpha-beta heterodimers of MHC-restricted T cell clones. Cell. 1986 Oct 24;47(2):161–171. doi: 10.1016/0092-8674(86)90439-3. [DOI] [PubMed] [Google Scholar]
  31. Siliciano R. F., Pratt J. C., Schmidt R. E., Ritz J., Reinherz E. L. Activation of cytolytic T lymphocyte and natural killer cell function through the T11 sheep erythrocyte binding protein. Nature. 1985 Oct 3;317(6036):428–430. doi: 10.1038/317428a0. [DOI] [PubMed] [Google Scholar]
  32. Springer T. A., Mann D. L., DeFranco A. L., Strominger J. L. Detergent solubilization, purification, and separation of specificities of HLA antigens from a cultured human lymphoblastoid line, RPMI 4265. J Biol Chem. 1977 Jul 10;252(13):4682–4693. [PubMed] [Google Scholar]
  33. Swain S. L. T cell subsets and the recognition of MHC class. Immunol Rev. 1983;74:129–142. doi: 10.1111/j.1600-065x.1983.tb01087.x. [DOI] [PubMed] [Google Scholar]
  34. Tadmori W., Reed J. C., Nowell P. C., Kamoun M. Functional properties of the 50 kd protein associated with the E-receptor on human T lymphocytes: suppression of IL 2 production by anti-p50 monoclonal antibodies. J Immunol. 1985 Mar;134(3):1709–1716. [PubMed] [Google Scholar]
  35. Tomonari K. Cytotoxic T cells generated in the autologous mixed lymphocyte reaction. I. Primary autologous mixed lymphocyte reaction. J Immunol. 1980 Mar;124(3):1111–1121. [PubMed] [Google Scholar]
  36. Van Wauwe J., Goossens J., Decock W., Kung P., Goldstein G. Suppression of human T-cell mitogenesis and E-rosette formation by the monoclonal antibody OKT11A. Immunology. 1981 Dec;44(4):865–871. [PMC free article] [PubMed] [Google Scholar]
  37. Watts T. H., Gaub H. E., McConnell H. M. T-cell-mediated association of peptide antigen and major histocompatibility complex protein detected by energy transfer in an evanescent wave-field. Nature. 1986 Mar 13;320(6058):179–181. doi: 10.1038/320179a0. [DOI] [PubMed] [Google Scholar]

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