Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1988 Dec;62(12):4452–4464. doi: 10.1128/jvi.62.12.4452-4464.1988

Soluble gp350/220 and deletion mutant glycoproteins block Epstein-Barr virus adsorption to lymphocytes.

J Tanner 1, Y Whang 1, J Sample 1, A Sears 1, E Kieff 1
PMCID: PMC253554  PMID: 2460635

Abstract

The Epstein-Barr virus (EBV) major outer envelope glycoprotein complex, gp350/220, was known to be a ligand for CR2, a B-lymphocyte plasma membrane protein. By Scatchard analysis, soluble EBV gp350/220 binds with high affinity (KD, 1.2 x 10(-8) M) to approximately the same number of B-lymphocyte surface sites as do CR2-specific monoclonal antibodies. Soluble gp350, gp220, or an amino-terminal, 576-amino-acid gp220 derivative binds similarly to B-lymphocyte receptors. Soluble gp350/220, gp220, or even a 470-amino-acid, amino-terminal gp220 derivative blocks EBV adsorption or infection. These experiments demonstrate that (i) gp350/220 is the predominant or exclusive EBV ligand for B lymphocytes; (ii) ligand-receptor blockade can prevent lymphocyte infection by EBV; and (iii) the amino-terminal, 470-amino-acid domain of gp350/220 contains the key ligand domain(s). Consistent with the ligand domain(s) being in the amino-terminal half of gp220 are the findings that the gp350/220-specific, EBV-neutralizing monoclonal antibody 72A1 blocks EBV adsorption by recognizing an epitope in the amino-terminal 470 (probably within the amino-terminal 162) amino acids and a deletion of amino-terminal amino acids 28 and 29 from gp350/220 inactivates ligand activity.

Full text

PDF
4452

Images in this article

Selected References

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

  1. Armstrong G. D., Paul R. W., Lee P. W. Studies on reovirus receptors of L cells: virus binding characteristics and comparison with reovirus receptors of erythrocytes. Virology. 1984 Oct 15;138(1):37–48. doi: 10.1016/0042-6822(84)90145-4. [DOI] [PubMed] [Google Scholar]
  2. Beisel C., Tanner J., Matsuo T., Thorley-Lawson D., Kezdy F., Kieff E. Two major outer envelope glycoproteins of Epstein-Barr virus are encoded by the same gene. J Virol. 1985 Jun;54(3):665–674. doi: 10.1128/jvi.54.3.665-674.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Biggin M., Farrell P. J., Barrell B. G. Transcription and DNA sequence of the BamHI L fragment of B95-8 Epstein-Barr virus. EMBO J. 1984 May;3(5):1083–1090. doi: 10.1002/j.1460-2075.1984.tb01933.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen J. H., Fischer E., Kazatchkine M. D., Brochier J., Revillard J. P. Characterization of monoclonal antihuman-B-cell antibody BL13 as an anti-C3d-receptor (CR2) antibody. Scand J Immunol. 1986 Mar;23(3):279–285. doi: 10.1111/j.1365-3083.1986.tb01969.x. [DOI] [PubMed] [Google Scholar]
  5. Deen K. C., McDougal J. S., Inacker R., Folena-Wasserman G., Arthos J., Rosenberg J., Maddon P. J., Axel R., Sweet R. W. A soluble form of CD4 (T4) protein inhibits AIDS virus infection. Nature. 1988 Jan 7;331(6151):82–84. doi: 10.1038/331082a0. [DOI] [PubMed] [Google Scholar]
  6. Dolyniuk M., Wolff E., Kieff E. Proteins of Epstein-Barr Virus. II. Electrophoretic analysis of the polypeptides of the nucleocapsid and the glucosamine- and polysaccharide-containing components of enveloped virus. J Virol. 1976 Apr;18(1):289–297. doi: 10.1128/jvi.18.1.289-297.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edson C. M., Thorley-Lawson D. A. Epstein-Barr virus membrane antigens: characterization, distribution, and strain differences. J Virol. 1981 Jul;39(1):172–184. doi: 10.1128/jvi.39.1.172-184.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edson C. M., Thorley-Lawson D. A. Synthesis and processing of the three major envelope glycoproteins of Epstein-Barr virus. J Virol. 1983 May;46(2):547–556. doi: 10.1128/jvi.46.2.547-556.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Einhorn L., Steinitz M., Yefenof E., Ernberg I., Bakacs T., Klein G. Epstein-Barr virus (EBV) receptors, complement receptors, and EBV infectibility of different lymphocyte fractions of human peripheral blood. II. Epstein-Barr virus studies. Cell Immunol. 1978 Jan;35(1):43–58. doi: 10.1016/0008-8749(78)90125-9. [DOI] [PubMed] [Google Scholar]
  10. Epstein M. A., Morgan A. J., Finerty S., Randle B. J., Kirkwood J. K. Protection of cottontop tamarins against Epstein-Barr virus-induced malignant lymphoma by a prototype subunit vaccine. Nature. 1985 Nov 21;318(6043):287–289. doi: 10.1038/318287a0. [DOI] [PubMed] [Google Scholar]
  11. Fingeroth J. D., Weis J. J., Tedder T. F., Strominger J. L., Biro P. A., Fearon D. T. Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4510–4514. doi: 10.1073/pnas.81.14.4510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fisher R. A., Bertonis J. M., Meier W., Johnson V. A., Costopoulos D. S., Liu T., Tizard R., Walker B. D., Hirsch M. S., Schooley R. T. HIV infection is blocked in vitro by recombinant soluble CD4. Nature. 1988 Jan 7;331(6151):76–78. doi: 10.1038/331076a0. [DOI] [PubMed] [Google Scholar]
  13. Fuller A. O., Spear P. G. Specificities of monoclonal and polyclonal antibodies that inhibit adsorption of herpes simplex virus to cells and lack of inhibition by potent neutralizing antibodies. J Virol. 1985 Aug;55(2):475–482. doi: 10.1128/jvi.55.2.475-482.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hinuma Y., Grace J. T., Jr Cloning of immunoglobulin-producing human leukemic and lymphoma cells in long-term cultures. Proc Soc Exp Biol Med. 1967 Jan;124(1):107–111. doi: 10.3181/00379727-124-31677. [DOI] [PubMed] [Google Scholar]
  15. Hoffman G. J., Lazarowitz S. G., Hayward S. D. Monoclonal antibody against a 250,000-dalton glycoprotein of Epstein-Barr virus identifies a membrane antigen and a neutralizing antigen. Proc Natl Acad Sci U S A. 1980 May;77(5):2979–2983. doi: 10.1073/pnas.77.5.2979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holland T. C., Homa F. L., Marlin S. D., Levine M., Glorioso J. Herpes simplex virus type 1 glycoprotein C-negative mutants exhibit multiple phenotypes, including secretion of truncated glycoproteins. J Virol. 1984 Nov;52(2):566–574. doi: 10.1128/jvi.52.2.566-574.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Houghten R. A. General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5131–5135. doi: 10.1073/pnas.82.15.5131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hummel M., Thorley-Lawson D., Kieff E. An Epstein-Barr virus DNA fragment encodes messages for the two major envelope glycoproteins (gp350/300 and gp220/200). J Virol. 1984 Feb;49(2):413–417. doi: 10.1128/jvi.49.2.413-417.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hussey R. E., Richardson N. E., Kowalski M., Brown N. R., Chang H. C., Siliciano R. F., Dorfman T., Walker B., Sodroski J., Reinherz E. L. A soluble CD4 protein selectively inhibits HIV replication and syncytium formation. Nature. 1988 Jan 7;331(6151):78–81. doi: 10.1038/331078a0. [DOI] [PubMed] [Google Scholar]
  20. Hutt-Fletcher L. M., Fowler E., Lambris J. D., Feighny R. J., Simmons J. G., Ross G. D. Studies of the Epstein Barr virus receptor found on Raji cells. II. A comparison of lymphocyte binding sites for Epstein Barr virus and C3d. J Immunol. 1983 Mar;130(3):1309–1312. [PubMed] [Google Scholar]
  21. Jondal M., Klein G., Oldstone M. B., Bokish V., Yefenof E. Surface markers on human B and T lymphocytes. VIII. Association between complement and Epstein-Barr virus receptors on human lymphoid cells. Scand J Immunol. 1976;5(4):401–410. doi: 10.1111/j.1365-3083.1976.tb00294.x. [DOI] [PubMed] [Google Scholar]
  22. Lambris J. D., Ganu V. S., Hirani S., Müller-Eberhard H. J. Mapping of the C3d receptor (CR2)-binding site and a neoantigenic site in the C3d domain of the third component of complement. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4235–4239. doi: 10.1073/pnas.82.12.4235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lasky L. A., Nakamura G., Smith D. H., Fennie C., Shimasaki C., Patzer E., Berman P., Gregory T., Capon D. J. Delineation of a region of the human immunodeficiency virus type 1 gp120 glycoprotein critical for interaction with the CD4 receptor. Cell. 1987 Sep 11;50(6):975–985. doi: 10.1016/0092-8674(87)90524-1. [DOI] [PubMed] [Google Scholar]
  24. Lischwe M. A., Ochs D. A new method for partial peptide mapping using N-chlorosuccinimide/urea and peptide silver staining in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1982 Dec;127(2):453–457. doi: 10.1016/0003-2697(82)90203-2. [DOI] [PubMed] [Google Scholar]
  25. Lowe R. S., Keller P. M., Keech B. J., Davison A. J., Whang Y., Morgan A. J., Kieff E., Ellis R. W. Varicella-zoster virus as a live vector for the expression of foreign genes. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3896–3900. doi: 10.1073/pnas.84.11.3896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mackett M., Arrand J. R. Recombinant vaccinia virus induces neutralising antibodies in rabbits against Epstein-Barr virus membrane antigen gp340. EMBO J. 1985 Dec 1;4(12):3229–3234. doi: 10.1002/j.1460-2075.1985.tb04070.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mann K. P., Staunton D., Thorley-Lawson D. A. Epstein-Barr virus-encoded protein found in plasma membranes of transformed cells. J Virol. 1985 Sep;55(3):710–720. doi: 10.1128/jvi.55.3.710-720.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McDougal J. S., Mawle A., Cort S. P., Nicholson J. K., Cross G. D., Scheppler-Campbell J. A., Hicks D., Sligh J. Cellular tropism of the human retrovirus HTLV-III/LAV. I. Role of T cell activation and expression of the T4 antigen. J Immunol. 1985 Nov;135(5):3151–3162. [PubMed] [Google Scholar]
  29. Miller G., Robinson J., Heston L., Lipman M. Differences between laboratory strains of Epstein-Barr virus based on immortalization, abortive infection, and interference. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4006–4010. doi: 10.1073/pnas.71.10.4006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moore M. D., Cooper N. R., Tack B. F., Nemerow G. R. Molecular cloning of the cDNA encoding the Epstein-Barr virus/C3d receptor (complement receptor type 2) of human B lymphocytes. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9194–9198. doi: 10.1073/pnas.84.24.9194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Nemerow G. R., Cooper N. R. Early events in the infection of human B lymphocytes by Epstein-Barr virus: the internalization process. Virology. 1984 Jan 15;132(1):186–198. doi: 10.1016/0042-6822(84)90102-8. [DOI] [PubMed] [Google Scholar]
  33. Nemerow G. R., Mold C., Schwend V. K., Tollefson V., Cooper N. R. Identification of gp350 as the viral glycoprotein mediating attachment of Epstein-Barr virus (EBV) to the EBV/C3d receptor of B cells: sequence homology of gp350 and C3 complement fragment C3d. J Virol. 1987 May;61(5):1416–1420. doi: 10.1128/jvi.61.5.1416-1420.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nemerow G. R., Siaw M. F., Cooper N. R. Purification of the Epstein-Barr virus/C3d complement receptor of human B lymphocytes: antigenic and functional properties of the purified protein. J Virol. 1986 May;58(2):709–712. doi: 10.1128/jvi.58.2.709-712.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nemerow G. R., Wolfert R., McNaughton M. E., Cooper N. R. Identification and characterization of the Epstein-Barr virus receptor on human B lymphocytes and its relationship to the C3d complement receptor (CR2). J Virol. 1985 Aug;55(2):347–351. doi: 10.1128/jvi.55.2.347-351.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nikodem V., Fresco J. R. Protein fingerprinting by SDS-gel electrophoresis after partial fragmentation with CNBr. Anal Biochem. 1979 Sep 1;97(2):382–386. doi: 10.1016/0003-2697(79)90089-7. [DOI] [PubMed] [Google Scholar]
  37. PULVERTAFT J. V. CYTOLOGY OF BURKITT'S TUMOUR (AFRICAN LYMPHOMA). Lancet. 1964 Feb 1;1(7327):238–240. doi: 10.1016/s0140-6736(64)92345-1. [DOI] [PubMed] [Google Scholar]
  38. Qualtiere L. F., Decoteau J. F., Hassan Nasr-el-Din M. Epitope mapping of the major Epstein-Barr virus outer envelope glycoprotein gp350/220. J Gen Virol. 1987 Feb;68(Pt 2):535–543. doi: 10.1099/0022-1317-68-2-535. [DOI] [PubMed] [Google Scholar]
  39. Rothman J. E. Protein sorting by selective retention in the endoplasmic reticulum and Golgi stack. Cell. 1987 Aug 14;50(4):521–522. doi: 10.1016/0092-8674(87)90024-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Royston I., Smith R. W., Buell D. N., Huang E. S., Pagano J. S. Autologous human B and T lymphoblastoid cell lines. Nature. 1974 Oct 25;251(5477):745–746. doi: 10.1038/251745a0. [DOI] [PubMed] [Google Scholar]
  41. Sixbey J. W., Davis D. S., Young L. S., Hutt-Fletcher L., Tedder T. F., Rickinson A. B. Human epithelial cell expression of an Epstein-Barr virus receptor. J Gen Virol. 1987 Mar;68(Pt 3):805–811. doi: 10.1099/0022-1317-68-3-805. [DOI] [PubMed] [Google Scholar]
  42. Smith D. H., Byrn R. A., Marsters S. A., Gregory T., Groopman J. E., Capon D. J. Blocking of HIV-1 infectivity by a soluble, secreted form of the CD4 antigen. Science. 1987 Dec 18;238(4834):1704–1707. doi: 10.1126/science.3500514. [DOI] [PubMed] [Google Scholar]
  43. Strnad B. C., Neubauer R. H., Rabin H., Mazur R. A. Correlation between Epstein-Barr virus membrane antigen and three large cell surface glycoproteins. J Virol. 1979 Dec;32(3):885–894. doi: 10.1128/jvi.32.3.885-894.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Strnad B. C., Schuster T., Klein R., Hopkins R. F., 3rd, Witmer T., Neubauer R. H., Rabin H. Production and characterization of monoclonal antibodies against the Epstein-Barr virus membrane antigen. J Virol. 1982 Jan;41(1):258–264. doi: 10.1128/jvi.41.1.258-264.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tanner J., Weis J., Fearon D., Whang Y., Kieff E. Epstein-Barr virus gp350/220 binding to the B lymphocyte C3d receptor mediates adsorption, capping, and endocytosis. Cell. 1987 Jul 17;50(2):203–213. doi: 10.1016/0092-8674(87)90216-9. [DOI] [PubMed] [Google Scholar]
  46. Tedder T. F., Clement L. T., Cooper M. D. Expression of C3d receptors during human B cell differentiation: immunofluorescence analysis with the HB-5 monoclonal antibody. J Immunol. 1984 Aug;133(2):678–683. [PubMed] [Google Scholar]
  47. Tedder T. F., Goldmacher V. S., Lambert J. M., Schlossman S. F. Epstein Barr virus binding induces internalization of the C3d receptor: a novel immunotoxin delivery system. J Immunol. 1986 Aug 15;137(4):1387–1391. [PubMed] [Google Scholar]
  48. Thorley-Lawson D. A., Edson C. M. Polypeptides of the Epstein-Barr virus membrane antigen complex. J Virol. 1979 Nov;32(2):458–467. doi: 10.1128/jvi.32.2.458-467.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Thorley-Lawson D. A., Poodry C. A. Identification and isolation of the main component (gp350-gp220) of Epstein-Barr virus responsible for generating neutralizing antibodies in vivo. J Virol. 1982 Aug;43(2):730–736. doi: 10.1128/jvi.43.2.730-736.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Weis J. J., Fearon D. T. The identification of N-linked oligosaccharides on the human CR2/Epstein-Barr virus receptor and their function in receptor metabolism, plasma membrane expression, and ligand binding. J Biol Chem. 1985 Nov 5;260(25):13824–13830. [PubMed] [Google Scholar]
  51. Weis J. J., Tedder T. F., Fearon D. T. Identification of a 145,000 Mr membrane protein as the C3d receptor (CR2) of human B lymphocytes. Proc Natl Acad Sci U S A. 1984 Feb;81(3):881–885. doi: 10.1073/pnas.81.3.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wells A., Koide N., Klein G. Two large virion envelope glycoproteins mediate Epstein-Barr virus binding to receptor-positive cells. J Virol. 1982 Jan;41(1):286–297. doi: 10.1128/jvi.41.1.286-297.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Whang Y., Silberklang M., Morgan A., Munshi S., Lenny A. B., Ellis R. W., Kieff E. Expression of the Epstein-Barr virus gp350/220 gene in rodent and primate cells. J Virol. 1987 Jun;61(6):1796–1807. doi: 10.1128/jvi.61.6.1796-1807.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Young L. S., Clark D., Sixbey J. W., Rickinson A. B. Epstein-Barr virus receptors on human pharyngeal epithelia. Lancet. 1986 Feb 1;1(8475):240–242. doi: 10.1016/s0140-6736(86)90776-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES