Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1991 Apr 1;173(4):971–980. doi: 10.1084/jem.173.4.971

Infection of human thymocytes by Epstein-Barr virus

PMCID: PMC2190801  PMID: 1706754

Abstract

The Epstein-Barr Virus (EBV) causes infectious mononucleosis, and has been strongly associated with certain human cancers. The virus is thought to exclusively bind to B lymphocytes and epithelial cells via receptors (CR2/CD21) that also interact with fragments of the third component of complement (C3). Recent evidence, however, has challenged this belief. We have used two-color immunofluorescence analysis using biotin-conjugated EBV and streptavidin-phycoerythrin along with fluorescein-conjugated anti-T cell antibodies and demonstrated that CD1- positive, CD3-dull (immature) human thymocytes express functional EBV receptors. In four replicate experiments, the binding of EBV to thymocytes ranged between 8 and 18%. This interaction is specific as evidenced by inhibition with nonconjugated virus, anti-CR2 antibodies, aggregated C3, and an antibody to the gp350 viral glycoprotein that the virus uses to bind to CR2. EBV can infect the thymocytes as evaluated by the presence of episomal EBV-DNA in thymocytes that had been incubated with the virus as short as 12 days or as long as 6 weeks. Episomal DNA analysis was performed by Southern blotting with a EBV-DNA probe that hybridizes to the first internal reiteration of the viral DNA. The presence of the EBV genome is also supported by the detection of EBV nuclear antigen 1 in infected thymocytes, assessed by Western blotting with EBV-immune sera. The EBV infection is specific as determined by blocking experiments using anti-CR2 and anti-gp350 antibodies. Finally, virus infection of thymocytes can act synergistically along with interleukin 2 and induce a lymphokine- dependent cellular proliferation. In view of previously reported cases of EBV-positive human T cell lymphomas, the possibility is raised that EBV may be involved in cancers of T lymphocytes that have not been previously appreciated.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Anderson D. L., Tsoukas C. D. Cholera toxin inhibits resting human T cell activation via a cAMP-independent pathway. J Immunol. 1989 Dec 1;143(11):3647–3652. [PubMed] [Google Scholar]
  2. Becherer J. D., Alsenz J., Servis C., Myones B. L., Lambris J. D. Cell surface proteins reacting with activated complement components. Complement Inflamm. 1989;6(3):142–165. doi: 10.1159/000463091. [DOI] [PubMed] [Google Scholar]
  3. Bergman Y., Stewart S. J., Levy S., Levy R. Biosynthesis, glycosylation, and in vitro translation of the human T cell antigen Leu-4. J Immunol. 1983 Oct;131(4):1876–1881. [PubMed] [Google Scholar]
  4. Carel J. C., Myones B. L., Frazier B., Holers V. M. Structural requirements for C3d,g/Epstein-Barr virus receptor (CR2/CD21) ligand binding, internalization, and viral infection. J Biol Chem. 1990 Jul 25;265(21):12293–12299. [PubMed] [Google Scholar]
  5. Cheung A., Kieff E. Long internal direct repeat in Epstein-Barr virus DNA. J Virol. 1982 Oct;44(1):286–294. doi: 10.1128/jvi.44.1.286-294.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Epstein M. A., Achong B. G. Recent progress in Epstein-Barr virus research. Annu Rev Microbiol. 1977;31:421–445. doi: 10.1146/annurev.mi.31.100177.002225. [DOI] [PubMed] [Google Scholar]
  7. Erdei A., Melchers F., Schulz T., Dierich M. The action of human C3 in soluble or cross-linked form with resting and activated murine B lymphocytes. Eur J Immunol. 1985 Feb;15(2):184–188. doi: 10.1002/eji.1830150214. [DOI] [PubMed] [Google Scholar]
  8. Fingeroth J. D., Clabby M. L., Strominger J. D. Characterization of a T-lymphocyte Epstein-Barr virus/C3d receptor (CD21). J Virol. 1988 Apr;62(4):1442–1447. doi: 10.1128/jvi.62.4.1442-1447.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Frade R., Barel M., Ehlin-Henriksson B., Klein G. gp140, the C3d receptor of human B lymphocytes, is also the Epstein-Barr virus receptor. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1490–1493. doi: 10.1073/pnas.82.5.1490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frade R., Crevon M. C., Barel M., Vazquez A., Krikorian L., Charriaut C., Galanaud P. Enhancement of human B cell proliferation by an antibody to the C3d receptor, the gp 140 molecule. Eur J Immunol. 1985 Jan;15(1):73–76. doi: 10.1002/eji.1830150114. [DOI] [PubMed] [Google Scholar]
  12. Fujisaku A., Harley J. B., Frank M. B., Gruner B. A., Frazier B., Holers V. M. Genomic organization and polymorphisms of the human C3d/Epstein-Barr virus receptor. J Biol Chem. 1989 Feb 5;264(4):2118–2125. [PubMed] [Google Scholar]
  13. Gilhus N. E., Frostad S., Matre R., Tönder O. The ontogeny of immunological receptors in the thymus. Acta Pathol Microbiol Immunol Scand C. 1982 Dec;90(6):307–314. doi: 10.1111/j.1699-0463.1982.tb01455.x. [DOI] [PubMed] [Google Scholar]
  14. Harabuchi Y., Yamanaka N., Kataura A., Imai S., Kinoshita T., Mizuno F., Osato T. Epstein-Barr virus in nasal T-cell lymphomas in patients with lethal midline granuloma. Lancet. 1990 Jan 20;335(8682):128–130. doi: 10.1016/0140-6736(90)90002-m. [DOI] [PubMed] [Google Scholar]
  15. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Jones J. F., Shurin S., Abramowsky C., Tubbs R. R., Sciotto C. G., Wahl R., Sands J., Gottman D., Katz B. Z., Sklar J. T-cell lymphomas containing Epstein-Barr viral DNA in patients with chronic Epstein-Barr virus infections. N Engl J Med. 1988 Mar 24;318(12):733–741. doi: 10.1056/NEJM198803243181203. [DOI] [PubMed] [Google Scholar]
  18. Kikuta H., Taguchi Y., Tomizawa K., Kojima K., Kawamura N., Ishizaka A., Sakiyama Y., Matsumoto S., Imai S., Kinoshita T. Epstein-Barr virus genome-positive T lymphocytes in a boy with chronic active EBV infection associated with Kawasaki-like disease. Nature. 1988 Jun 2;333(6172):455–457. doi: 10.1038/333455a0. [DOI] [PubMed] [Google Scholar]
  19. Lambris J. D., Dobson N. J., Ross G. D. Release of endogenous C3b inactivator from lymphocytes in response to triggering membrane receptors for beta 1H globulin. J Exp Med. 1980 Dec 1;152(6):1625–1644. doi: 10.1084/jem.152.6.1625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lowell C. A., Klickstein L. B., Carter R. H., Mitchell J. A., Fearon D. T., Ahearn J. M. Mapping of the Epstein-Barr virus and C3dg binding sites to a common domain on complement receptor type 2. J Exp Med. 1989 Dec 1;170(6):1931–1946. doi: 10.1084/jem.170.6.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meeker T. C., Miller R. A., Link M. P., Bindl J., Warnke R., Levy R. A unique human B lymphocyte antigen defined by a monoclonal antibody. Hybridoma. 1984 Winter;3(4):305–320. doi: 10.1089/hyb.1984.3.305. [DOI] [PubMed] [Google Scholar]
  22. Melchers F., Erdei A., Schulz T., Dierich M. P. Growth control of activated, synchronized murine B cells by the C3d fragment of human complement. Nature. 1985 Sep 19;317(6034):264–267. doi: 10.1038/317264a0. [DOI] [PubMed] [Google Scholar]
  23. Menezes J., Seigneurin J. M., Patel P., Bourkas A., Lenoir G. Presence of Epstein-Barr virus receptors, but absence of virus penetration, in cells of an Epstein-Barr virus genome-negative human lymphoblastoid T line (Molt 4). J Virol. 1977 Jun;22(3):816–821. doi: 10.1128/jvi.22.3.816-821.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Middeldorp J. M., Herbrink P. Epstein-Barr virus specific marker molecules for early diagnosis of infectious mononucleosis. J Virol Methods. 1988 Sep;21(1-4):133–146. doi: 10.1016/0166-0934(88)90060-2. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  29. Reinherz E. L., Schlossman S. F. The differentiation and function of human T lymphocytes. Cell. 1980 Apr;19(4):821–827. doi: 10.1016/0092-8674(80)90072-0. [DOI] [PubMed] [Google Scholar]
  30. Reynes M., Aubert J. P., Cohen J. H., Audouin J., Tricottet V., Diebold J., Kazatchkine M. D. Human follicular dendritic cells express CR1, CR2, and CR3 complement receptor antigens. J Immunol. 1985 Oct;135(4):2687–2694. [PubMed] [Google Scholar]
  31. Ross G. D., Medof M. E. Membrane complement receptors specific for bound fragments of C3. Adv Immunol. 1985;37:217–267. doi: 10.1016/s0065-2776(08)60341-7. [DOI] [PubMed] [Google Scholar]
  32. Rumpold H., Rhodes G. H., Bloch P. L., Carson D. A., Vaughan J. H. The glycine-alanine repeating region is the major epitope of the Epstein-Barr nuclear antigen-1 (EBNA-1). J Immunol. 1987 Jan 15;138(2):593–599. [PubMed] [Google Scholar]
  33. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  34. Sauvageau G., Stocco R., Kasparian S., Menezes J. Epstein-Barr virus receptor expression on human CD8+ (cytotoxic/suppressor) T lymphocytes. J Gen Virol. 1990 Feb;71(Pt 2):379–386. doi: 10.1099/0022-1317-71-2-379. [DOI] [PubMed] [Google Scholar]
  35. Schreiber R. D. The chemistry and biology of complement receptors. Springer Semin Immunopathol. 1984;7(2-3):221–249. doi: 10.1007/BF01893021. [DOI] [PubMed] [Google Scholar]
  36. Servis C., Lambris J. D. C3 synthetic peptides support growth of human CR2-positive lymphoblastoid B cells. J Immunol. 1989 Apr 1;142(7):2207–2212. [PubMed] [Google Scholar]
  37. Shapiro I. M., Volsky D. J., Saemundsen A. K., Anisimova E., Klein G. Infection of the human T-cell-derived leukemia line Molt-4 by Epstein-Barr virus (EBV): induction of EBV-determined antigens and virus reproduction. Virology. 1982 Jul 15;120(1):171–181. doi: 10.1016/0042-6822(82)90015-0. [DOI] [PubMed] [Google Scholar]
  38. Shevach E., Edelson R., Frank M., Lutzner M., Green I. A human leukemia cell with both B and T cell surface receptors. Proc Natl Acad Sci U S A. 1974 Mar;71(3):863–866. doi: 10.1073/pnas.71.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shore A., Dosch H. M., Gelfand E. W. Expression and modulation of C3 receptors during early T-cell ontogeny. Cell Immunol. 1979 Jun;45(1):157–166. doi: 10.1016/0008-8749(79)90371-x. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. Tanner J., Whang Y., Sample J., Sears A., Kieff E. Soluble gp350/220 and deletion mutant glycoproteins block Epstein-Barr virus adsorption to lymphocytes. J Virol. 1988 Dec;62(12):4452–4464. doi: 10.1128/jvi.62.12.4452-4464.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Toben H. R., Smith R. G. T lymphocytes bearing complement receptors in a patient with chronic lymphocytic leukaemia. Clin Exp Immunol. 1977 Feb;27(2):292–302. [PMC free article] [PubMed] [Google Scholar]
  45. Tsoukas C. D., Lambris J. D. Expression of CR2/EBV receptors on human thymocytes detected by monoclonal antibodies. Eur J Immunol. 1988 Aug;18(8):1299–1302. doi: 10.1002/eji.1830180823. [DOI] [PubMed] [Google Scholar]
  46. Tsoukas C. D., Landgraf B., Bentin J., Lamberti J. F., Carson D. A., Vaughan J. H. Structural and functional characteristics of the CD3 (T3) molecular complex on human thymocytes. J Immunol. 1987 Jun 1;138(11):3885–3890. [PubMed] [Google Scholar]
  47. Vik D. P., Fearon D. T. Cellular distribution of complement receptor type 4 (CR4): expression on human platelets. J Immunol. 1987 Jan 1;138(1):254–258. [PubMed] [Google Scholar]
  48. Wahl G. M., Meinkoth J. L., Kimmel A. R. Northern and Southern blots. Methods Enzymol. 1987;152:572–581. doi: 10.1016/0076-6879(87)52064-x. [DOI] [PubMed] [Google Scholar]
  49. Wang A., Lu S. D., Mark D. F. Site-specific mutagenesis of the human interleukin-2 gene: structure-function analysis of the cysteine residues. Science. 1984 Jun 29;224(4656):1431–1433. doi: 10.1126/science.6427925. [DOI] [PubMed] [Google Scholar]
  50. Weis J. J., Toothaker L. E., Smith J. A., Weis J. H., Fearon D. T. Structure of the human B lymphocyte receptor for C3d and the Epstein-Barr virus and relatedness to other members of the family of C3/C4 binding proteins. J Exp Med. 1988 Mar 1;167(3):1047–1066. doi: 10.1084/jem.167.3.1047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. West W., Herberman R. B. A human lymphoid cell line with receptors for both sheep red blood cells and complement. Cell Immunol. 1974 Oct;14(1):139–145. doi: 10.1016/0008-8749(74)90178-6. [DOI] [PubMed] [Google Scholar]
  52. Witsell A. L., Schook L. B. Clonal analysis of gene expression by PCR. Biotechniques. 1990 Sep;9(3):318–322. [PubMed] [Google Scholar]
  53. Yates J. L., Warren N., Sugden B. Stable replication of plasmids derived from Epstein-Barr virus in various mammalian cells. 1985 Feb 28-Mar 6Nature. 313(6005):812–815. doi: 10.1038/313812a0. [DOI] [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 The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES