Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1993 May 1;177(5):1439–1450. doi: 10.1084/jem.177.5.1439

A soluble multimeric recombinant CD2 protein identifies CD48 as a low affinity ligand for human CD2: divergence of CD2 ligands during the evolution of humans and mice

PMCID: PMC2191021  PMID: 7683037

Abstract

To search for possible ligands of CD2 distinct from CD58 (lymphocyte function-associated antigen 3), we have produced a soluble pentameric CD2-immunoglobulin (Ig) fusion protein (spCD2) linking the 182-amino acid human CD2 extracellular segment with CH2-CH3-CH4 domains of human IgM heavy chain, thus enhancing the micromolar affinity of the CD2 monomer through multimeric interaction. Using quantitative immunofluorescence and standard stringency wash conditions, we observed that the binding of spCD2 to human B lymphoblastoid JY cells and red blood cells is virtually inhibited by anti-CD58 TS2/9 monoclonal antibody, even though these cells express levels of CD48 and CD59 comparable to CD58. Consistent with these results, spCD2 did not show any binding to Chinese hamster ovary (CHO) cells transfected with human CD48 or CD59. However, binding studies on CD48-, CD58-, or CD59- transfected CHO cells with spCD2 under low stringency wash conditions revealed that human CD48 is a low affinity ligand of human CD2 compared with CD58 (Kd approximately 10(-4) vs. approximately 10(-6) M, respectively). The findings are noteworthy given that in the murine system CD48 is the major ligand for CD2. No detectable binding was observed to CD59-transfected CHO cells despite a report suggesting that CD59 may bind to the human CD2 adhesion domain. Importantly, in cell- cell adhesion assays between CD2+ Jurkat T cells and CD48- or CD59- transfected CHO cells, there was no conjugate formation, whereas binding of Jurkat T cells to CD58-transfected CHO cells was readily detected. Collectively, our findings provide evidence for a conservation of the CD2-CD48 interaction in the human species that may be of limited, if any, functional significance. Given the importance of the CD2-CD48 interaction in the murine system and CD2-CD58 interaction in humans, it would appear that there has been a divergence of functional CD2 ligands during the evolution of humans and mice.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alcover A., Alberini C., Acuto O., Clayton L. K., Transy C., Spagnoli G. C., Moingeon P., Lopez P., Reinherz E. L. Interdependence of CD3-Ti and CD2 activation pathways in human T lymphocytes. EMBO J. 1988 Jul;7(7):1973–1977. doi: 10.1002/j.1460-2075.1988.tb03035.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bierer B. E., Sleckman B. P., Ratnofsky S. E., Burakoff S. J. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989;7:579–599. doi: 10.1146/annurev.iy.07.040189.003051. [DOI] [PubMed] [Google Scholar]
  3. Boulianne G. L., Hozumi N., Shulman M. J. Production of functional chimaeric mouse/human antibody. Nature. 1984 Dec 13;312(5995):643–646. doi: 10.1038/312643a0. [DOI] [PubMed] [Google Scholar]
  4. Clayton L. K., Sayre P. H., Novotny J., Reinherz E. L. Murine and human T11 (CD2) cDNA sequences suggest a common signal transduction mechanism. Eur J Immunol. 1987 Sep;17(9):1367–1370. doi: 10.1002/eji.1830170922. [DOI] [PubMed] [Google Scholar]
  5. Davies A., Simmons D. L., Hale G., Harrison R. A., Tighe H., Lachmann P. J., Waldmann H. CD59, an LY-6-like protein expressed in human lymphoid cells, regulates the action of the complement membrane attack complex on homologous cells. J Exp Med. 1989 Sep 1;170(3):637–654. doi: 10.1084/jem.170.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deckert M., Kubar J., Bernard A. CD58 and CD59 molecules exhibit potentializing effects in T cell adhesion and activation. J Immunol. 1992 Feb 1;148(3):672–677. [PubMed] [Google Scholar]
  7. Deckert M., Kubar J., Zoccola D., Bernard-Pomier G., Angelisova P., Horejsi V., Bernard A. CD59 molecule: a second ligand for CD2 in T cell adhesion. Eur J Immunol. 1992 Nov;22(11):2943–2947. doi: 10.1002/eji.1830221128. [DOI] [PubMed] [Google Scholar]
  8. Diamond M. S., Staunton D. E., Marlin S. D., Springer T. A. Binding of the integrin Mac-1 (CD11b/CD18) to the third immunoglobulin-like domain of ICAM-1 (CD54) and its regulation by glycosylation. Cell. 1991 Jun 14;65(6):961–971. doi: 10.1016/0092-8674(91)90548-d. [DOI] [PubMed] [Google Scholar]
  9. Doyle C., Strominger J. L. Interaction between CD4 and class II MHC molecules mediates cell adhesion. Nature. 1987 Nov 19;330(6145):256–259. doi: 10.1038/330256a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Freedman A. S., Freeman G., Horowitz J. C., Daley J., Nadler L. M. B7, a B-cell-restricted antigen that identifies preactivated B cells. J Immunol. 1987 Nov 15;139(10):3260–3267. [PubMed] [Google Scholar]
  12. Gay D., Buus S., Pasternak J., Kappler J., Marrack P. The T-cell accessory molecule CD4 recognizes a monomorphic determinant on isolated Ia. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5629–5633. doi: 10.1073/pnas.85.15.5629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gimmi C. D., Freeman G. J., Gribben J. G., Sugita K., Freedman A. S., Morimoto C., Nadler L. M. B-cell surface antigen B7 provides a costimulatory signal that induces T cells to proliferate and secrete interleukin 2. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6575–6579. doi: 10.1073/pnas.88.15.6575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Groux H., Huet S., Aubrit F., Tran H. C., Boumsell L., Bernard A. A 19-kDa human erythrocyte molecule H19 is involved in rosettes, present on nucleated cells, and required for T cell activation. Comparison of the roles of H19 and LFA-3 molecules in T cell activation. J Immunol. 1989 May 1;142(9):3013–3020. [PubMed] [Google Scholar]
  15. Hahn W. C., Menu E., Bothwell A. L., Sims P. J., Bierer B. E. Overlapping but nonidentical binding sites on CD2 for CD58 and a second ligand CD59. Science. 1992 Jun 26;256(5065):1805–1807. doi: 10.1126/science.1377404. [DOI] [PubMed] [Google Scholar]
  16. Hünig T., Tiefenthaler G., Meyer zum Büschenfelde K. H., Meuer S. C. Alternative pathway activation of T cells by binding of CD2 to its cell-surface ligand. Nature. 1987 Mar 19;326(6110):298–301. doi: 10.1038/326298a0. [DOI] [PubMed] [Google Scholar]
  17. Kato K., Koyanagi M., Okada H., Takanashi T., Wong Y. W., Williams A. F., Okumura K., Yagita H. CD48 is a counter-receptor for mouse CD2 and is involved in T cell activation. J Exp Med. 1992 Nov 1;176(5):1241–1249. doi: 10.1084/jem.176.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Killeen N., Stuart S. G., Littman D. R. Development and function of T cells in mice with a disrupted CD2 gene. EMBO J. 1992 Dec;11(12):4329–4336. doi: 10.1002/j.1460-2075.1992.tb05532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koyasu S., Lawton T., Novick D., Recny M. A., Siliciano R. F., Wallner B. P., Reinherz E. L. Role of interaction of CD2 molecules with lymphocyte function-associated antigen 3 in T-cell recognition of nominal antigen. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2603–2607. doi: 10.1073/pnas.87.7.2603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Linsley P. S., Brady W., Urnes M., Grosmaire L. S., Damle N. K., Ledbetter J. A. CTLA-4 is a second receptor for the B cell activation antigen B7. J Exp Med. 1991 Sep 1;174(3):561–569. doi: 10.1084/jem.174.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Linsley P. S., Clark E. A., Ledbetter J. A. T-cell antigen CD28 mediates adhesion with B cells by interacting with activation antigen B7/BB-1. Proc Natl Acad Sci U S A. 1990 Jul;87(13):5031–5035. doi: 10.1073/pnas.87.13.5031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Luo W., Van de Velde H., von Hoegen I., Parnes J. R., Thielemans K. Ly-1 (CD5), a membrane glycoprotein of mouse T lymphocytes and a subset of B cells, is a natural ligand of the B cell surface protein Lyb-2 (CD72). J Immunol. 1992 Mar 15;148(6):1630–1634. [PubMed] [Google Scholar]
  24. Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Moingeon P. E., Lucich J. L., Stebbins C. C., Recny M. A., Wallner B. P., Koyasu S., Reinherz E. L. Complementary roles for CD2 and LFA-1 adhesion pathways during T cell activation. Eur J Immunol. 1991 Mar;21(3):605–610. doi: 10.1002/eji.1830210311. [DOI] [PubMed] [Google Scholar]
  27. Moingeon P., Chang H. C., Sayre P. H., Clayton L. K., Alcover A., Gardner P., Reinherz E. L. The structural biology of CD2. Immunol Rev. 1989 Oct;111:111–144. doi: 10.1111/j.1600-065x.1989.tb00544.x. [DOI] [PubMed] [Google Scholar]
  28. Moingeon P., Chang H. C., Wallner B. P., Stebbins C., Frey A. Z., Reinherz E. L. CD2-mediated adhesion facilitates T lymphocyte antigen recognition function. Nature. 1989 May 25;339(6222):312–314. doi: 10.1038/339312a0. [DOI] [PubMed] [Google Scholar]
  29. Moseley W. S., Seldin M. F. Definition of mouse chromosome 1 and 3 gene linkage groups that are conserved on human chromosome 1: evidence that a conserved linkage group spans the centromere of human chromosome 1. Genomics. 1989 Nov;5(4):899–905. doi: 10.1016/0888-7543(89)90132-8. [DOI] [PubMed] [Google Scholar]
  30. Parish C. R., McPhun V., Warren H. S. Is a natural ligand of the T lymphocyte CD2 molecule a sulfated carbohydrate? J Immunol. 1988 Nov 15;141(10):3498–3504. [PubMed] [Google Scholar]
  31. Recny M. A., Luther M. A., Knoppers M. H., Neidhardt E. A., Khandekar S. S., Concino M. F., Schimke P. A., Francis M. A., Moebius U., Reinhold B. B. N-glycosylation is required for human CD2 immunoadhesion functions. J Biol Chem. 1992 Nov 5;267(31):22428–22434. [PubMed] [Google Scholar]
  32. 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]
  33. Sayre P. H., Hussey R. E., Chang H. C., Ciardelli T. L., Reinherz E. L. Structural and binding analysis of a two domain extracellular CD2 molecule. J Exp Med. 1989 Mar 1;169(3):995–1009. doi: 10.1084/jem.169.3.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. 1987 Oct 29-Nov 4Nature. 329(6142):840–842. doi: 10.1038/329840a0. [DOI] [PubMed] [Google Scholar]
  35. Selvaraj P., Plunkett M. L., Dustin M., Sanders M. E., Shaw S., Springer T. A. The T lymphocyte glycoprotein CD2 binds the cell surface ligand LFA-3. 1987 Mar 26-Apr 1Nature. 326(6111):400–403. doi: 10.1038/326400a0. [DOI] [PubMed] [Google Scholar]
  36. Sewell W. A., Palmer R. W., Spurr N. K., Sheer D., Brown M. H., Bell Y., Crumpton M. J. The human LFA-3 gene is located at the same chromosome band as the gene for its receptor CD2. Immunogenetics. 1988;28(4):278–282. doi: 10.1007/BF00345506. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Siu G., Hedrick S. M., Brian A. A. Isolation of the murine intercellular adhesion molecule 1 (ICAM-1) gene. ICAM-1 enhances antigen-specific T cell activation. J Immunol. 1989 Dec 1;143(11):3813–3820. [PubMed] [Google Scholar]
  39. Springer T. A., Dustin M. L., Kishimoto T. K., Marlin S. D. The lymphocyte function-associated LFA-1, CD2, and LFA-3 molecules: cell adhesion receptors of the immune system. Annu Rev Immunol. 1987;5:223–252. doi: 10.1146/annurev.iy.05.040187.001255. [DOI] [PubMed] [Google Scholar]
  40. Staunton D. E., Fisher R. C., LeBeau M. M., Lawrence J. B., Barton D. E., Francke U., Dustin M., Thorley-Lawson D. A. Blast-1 possesses a glycosyl-phosphatidylinositol (GPI) membrane anchor, is related to LFA-3 and OX-45, and maps to chromosome 1q21-23. J Exp Med. 1989 Mar 1;169(3):1087–1099. doi: 10.1084/jem.169.3.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Staunton D. E., Thorley-Lawson D. A. Molecular cloning of the lymphocyte activation marker Blast-1. EMBO J. 1987 Dec 1;6(12):3695–3701. doi: 10.1002/j.1460-2075.1987.tb02703.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Streuli M., Saito H. Regulation of tissue-specific alternative splicing: exon-specific cis-elements govern the splicing of leukocyte common antigen pre-mRNA. EMBO J. 1989 Mar;8(3):787–796. doi: 10.1002/j.1460-2075.1989.tb03439.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Subramani S., Mulligan R., Berg P. Expression of the mouse dihydrofolate reductase complementary deoxyribonucleic acid in simian virus 40 vectors. Mol Cell Biol. 1981 Sep;1(9):854–864. doi: 10.1128/mcb.1.9.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Van de Velde H., von Hoegen I., Luo W., Parnes J. R., Thielemans K. The B-cell surface protein CD72/Lyb-2 is the ligand for CD5. Nature. 1991 Jun 20;351(6328):662–665. doi: 10.1038/351662a0. [DOI] [PubMed] [Google Scholar]
  45. Vollger L. W., Tuck D. T., Springer T. A., Haynes B. F., Singer K. H. Thymocyte binding to human thymic epithelial cells is inhibited by monoclonal antibodies to CD-2 and LFA-3 antigens. J Immunol. 1987 Jan 15;138(2):358–363. [PubMed] [Google Scholar]
  46. Williams A. F., Barclay A. N. The immunoglobulin superfamily--domains for cell surface recognition. Annu Rev Immunol. 1988;6:381–405. doi: 10.1146/annurev.iy.06.040188.002121. [DOI] [PubMed] [Google Scholar]
  47. Wong Y. W., Williams A. F., Kingsmore S. F., Seldin M. F. Structure, expression, and genetic linkage of the mouse BCM1 (OX45 or Blast-1) antigen. Evidence for genetic duplication giving rise to the BCM1 region on mouse chromosome 1 and the CD2/LFA3 region on mouse chromosome 3. J Exp Med. 1990 Jun 1;171(6):2115–2130. doi: 10.1084/jem.171.6.2115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yokoyama S., Staunton D., Fisher R., Amiot M., Fortin J. J., Thorley-Lawson D. A. Expression of the Blast-1 activation/adhesion molecule and its identification as CD48. J Immunol. 1991 Apr 1;146(7):2192–2200. [PubMed] [Google Scholar]
  49. de Fougerolles A. R., Springer T. A. Intercellular adhesion molecule 3, a third adhesion counter-receptor for lymphocyte function-associated molecule 1 on resting lymphocytes. J Exp Med. 1992 Jan 1;175(1):185–190. doi: 10.1084/jem.175.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES