Expression of Virus-Associated Functions in Cells Transformed in Vitro by Epstein-Barr Virus: Epstein-Barr Virus Cell Surface Antigen and Virus-Release from Transformed Cells

  • Bill Sugden


Epstein-Barr virus (EBV) infects resting human B lymphocytes in vitro and induces up to 10% of the infected cells to proliferate as measured in a clonal transformation assay (1–5). This virus also infects B lymphocytes in vivo; up to 20% of the peripheral B cells in patients with infectious mononucleosis (IM) express an EBV-associated nuclear antigen, EBNA (6), When peripheral lymphocytes from IM patients are collected and cloned directly in soft agar, some B cells are found to proliferate indefinitely and to express EBNA (7). Although it is not known that virus infections in vitro and in vivo are functionally equivalent, no differences between B cells infected in vitro and in vivo have yet been identified once the cells are studied in vitro. We have studied cells transformed by EBV in vitro with the hope that such studies will help to define characteristics of some of the cells infected by EBV in vivo. The advantage of studying cells transformed in vitro is that we can chart their history from their exposure to EBV to the time of study.


Cell Surface Antigen Infectious Mononucleosis Seronegative Donor 2xl04 Cell National Cancer Institute Grant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Pope, J.H., Home, M.K., and Scott, W. Transformation of foetal human leukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int. J. Cancer, 3:857, 1968.PubMedCrossRefGoogle Scholar
  2. 2.
    Pattengale, P.K., Smith, R.W., and Gerber, P. Selective transformation of B lymphocytes by EB virus. Lancet, ii:93, 1973.CrossRefGoogle Scholar
  3. 3.
    Yamamoto, N. and Hinuma, Y. Clonal transformation of human leukocytes by Epstein-Barr virus in soft agar. Int. J. Cancer, 17:191, 1976.PubMedCrossRefGoogle Scholar
  4. 4.
    Sugden, B. and Mark, W. Clonal transformation of adult human leukocytes by Epstein-Barr virus. J. Virol., 23:503, 1977.PubMedGoogle Scholar
  5. 5.
    Henderson, E., Miller, G., Robinson, J., and Heston, L. Efficiency of transformation of lymphocytes by Epstein-Barr virus. Virology, 76:152, 1977.PubMedCrossRefGoogle Scholar
  6. 6.
    Robinson, J.E., Smith, D., and Niederraan, J. Plasmacytic differentiation of circulating Epstein-Barr virus-infected B lymphocytes during acute infectious mononucleosis. J. Exp. Med., 153:235, 1981.PubMedCrossRefGoogle Scholar
  7. 7.
    Hinuma, Y. and Katsuki, T. Colonies of EBNA-positive cells in soft agar from peripheral leukocytes of infectious mononucleosis patients. Int. J. Cancer, 21:426, 1978.PubMedCrossRefGoogle Scholar
  8. 8.
    Reedman, B.M. and Klein, G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int. J. Cancer, 11:499, 1973.PubMedCrossRefGoogle Scholar
  9. 9.
    van Santen, V., Cheung, A., and Kieff, E. Epstein-Barr virus RNA. VII. Size and direction of transcription of virus-specific cytoplasmic RNAs in a transformed cell line. Proc. Natl. Acad. Sci. USA, 78:1930, 1981.PubMedCrossRefGoogle Scholar
  10. 10.
    Kintner, C. and Sugden, B. Conservation and progressive methylation of Epstein-Barr virus DNA sequences in transformed cells. J. Virol., 38:305, 1981.PubMedGoogle Scholar
  11. 11.
    Lerner, M.R., Andrews, N.C., Miller, G., and Steitz, J.A. Two small RNAs encoded by Epstein-Barr virus and complexed with protein are precipitated by antibodies from patients with systemic lupus erythematosus. Proc. Natl. Acad. Sci. USA, 78: 805, 1981.PubMedCrossRefGoogle Scholar
  12. 12.
    Svedmyr, E. and Jondal, M. Cytotoxic effector cells specific for B cell lines transformed by Epstein-Barr virus are present in patients with infectious mononucleosis. Proc. Natl. Acad. Sci. USA, 72:1622, 1975.PubMedCrossRefGoogle Scholar
  13. 13.
    Royston, I., Sullivan, J.L., Periman, P.O., and Perlin, E. Cell-mediated immunity to Epstein-Barr-virus-transformed lymphoblastoid cells in acute infectious mononucleosis. N. Engl. J. Med., 293:1159, 1975.PubMedCrossRefGoogle Scholar
  14. 14.
    Svedmyr, E., Jondal, M., Henle, W., Weiland, O., Rorabo, L., and Klein, G. EBV-specific killer T cells and serologic responses after onset of infectious mononucleosis. J. Clin. Lab. Immunol., 1:225, 1978.PubMedGoogle Scholar
  15. 15.
    Sugamura, K. and Hinuma, Y. In vitro induction of cytotoxic T lymphocytes specific for Epstein-Barr virus-transformed cells: kinetics of autologous restimulation. J. Immunol., 124:1045, 1980.PubMedGoogle Scholar
  16. 16.
    Tanaka, Y., Sugamura, K., Hinuma, Y., Sato, H., and Okochi, K. Memory of Epstein-Barr virus-specific cytotoxic T cells in normal seropositive adults as revealed by an in vitro restimulation method. J. Immunol., 125:1426, 1980.”PubMedGoogle Scholar
  17. 17.
    Misko, I.S., Moss, D.J., and Pope, J.H. HLA antigen-related restriction of T lymphocyte cytotoxicity to Epstein-Barr virus. Proc. Natl. Acad. Sci. USA, 77:4247, 1980.PubMedCrossRefGoogle Scholar
  18. 18.
    Tsoukas, C.D., Fox, R.I., Slovin, S.F., Carson, D.A., Pellegrino, M., Fong, S., Pasquali, J.-L., Ferrone, S., Kung, P., and Vaughan, J.H. T lymphocyte-mediated cytotoxicity against autologous EBV-genome-bearing B cells. J. Immunol., 126:1742, 1981.PubMedGoogle Scholar
  19. 19.
    Rickinson, A.B., Moss, D.J., Allen, D.J., Wallace, L.E., Rowe, M., and Epstein, M.A. Reactivation of Epstein-Barr virus-specific cytotoxic T cells by in vitro stimulation with autologous lymphoblastoid cell line. Int. J. Cancer, 27:593, 1981.PubMedCrossRefGoogle Scholar
  20. 20.
    Lipinski, M., Fridman, W.H., Tursz, T., Vincent, C., Pious, D., and Fellous, M. Absence of allogeneic restriction in human T-cell-mediated cytotoxicity to Epstein-Barr virus-infected target cells. J. Exp. Med., 150:1310, 1979.PubMedCrossRefGoogle Scholar
  21. 21.
    Kintner, C. and Sugden, B. Identification of determinants unique to the surfaces of cells transformed by Epstein-Barr virus. Nature, 294:458, 1981.PubMedCrossRefGoogle Scholar
  22. 22.
    Slovin, S.F., Frisman, D.M., Tsoukas, C.D., Royston, I., Baird, S.M., Wormsley, S.B., Carson, D.A., and Vaughan, J.H. Membrane antigen on Epstein-Barr virus-infected human B cells recognized by a monoclonal antibody. Proc. Natl. Acad. Sci. USA, 79:2649, 1982.PubMedCrossRefGoogle Scholar
  23. 23.
    Rowe, M., Hildreth, J.E.K., Rickinson, A.B., and Epstein, M.A. Monoclonal antibodies to Epstein-Barr virus-induced, transformation-associated cell surface antigens: binding patterns and effect upon virus-specific T cell cytotoxicity. Int. J. Cancer, 29:373, 1982.PubMedCrossRefGoogle Scholar
  24. 24.
    Thorley-Lawson, D.A., Edson, C.M., and Geilinger, K. Epstein-Barr virus antigens — a challenge to modern biochemistry. In: G. Klein and S. Weinhouse (eds.), Advances in Cancer Research. Vol. 36, pp. 295–348. New York: Academic Press, 1982.Google Scholar
  25. 25.
    Sugden, B., Phelps, M., and Domoradzki, J. Epstein-Barr virus DNA is amplified in transformed lymphocytes. J. Virol., 31: 590, 1979.PubMedGoogle Scholar
  26. 26.
    Wilson, G. and Miller, G. Recovery of Epstein-Barr virus from nonproducer neonatal human lymphoid cell transformants. Virology, 95:351, 1979.PubMedCrossRefGoogle Scholar
  27. 26a.
    Thorley-Lawson, D.A., Schooley, R.T., Bhan, A.K., and Nadler, L.M. Epstein-Barr virus superinduces a new human B cell differentiation antigen (B-last-1) expressed on transformed cells. Cell, 30:415, 1982.PubMedCrossRefGoogle Scholar
  28. 27.
    Fu, S.M. and Hurley, J.N. Human cell lines containing Epstein-Barr virus but distinct from the common B cell lymphoblastoid lines. Proc. Natl. Acad. Sci. USA, 76:6637, 1979.PubMedCrossRefGoogle Scholar
  29. 28.
    Galfre, G. and Milstein, C. Preparation of monoclonal antibodies: strategies and procedures. In: J.J. Langone and H. van Vunakis (eds.), Method in Enzymology. Vol. 73, pp. 3–46. New York: Academic Press, 1981.Google Scholar
  30. 28a.
    Sugden, B. and Metzenberg, S. Characterization of an antigen whose cell surface expression is induced by infection with Epstein-Barr virus. J. Virol., 46:800, 1983.PubMedGoogle Scholar
  31. 29.
    Parham, P., Barnstable, C.J., and Bodmer, W.F. Use of monoclonal antibody (W6/32) in structural studies of HLA-A,B,C antigens. J. Immunol., 123: 342, 1979.PubMedGoogle Scholar
  32. 30.
    Strominger, J.L., Mann, D.L., Parham, P., Robb, R., Springer, T., and Terhorst, C. Structure of HLA-A and B antigens isolated from cultured human lymphocytes. Cold Spring Harbor Symp. Quant. Biol., 41:323, 1977.CrossRefGoogle Scholar
  33. 31.
    Brodsky, F.M., Parham, P., Barnstable, C.J., Crumpton, M.J., and Bodmer, W.F. Monoclonal antibodies for analysis of the HLA system. Immunological Rev., 47:3, 1979.CrossRefGoogle Scholar
  34. 32.
    Olsnes, S., Fernandez-Puentes, C., Carrasco, L., and Vasques, D. Ribosome inactivation by the toxic lectins abrin and ricin. Kinetics of the enzymic activity of the toxic A-chains. Eur. J. Biochem., 60:281, 1975.PubMedCrossRefGoogle Scholar
  35. 33.
    Cawley, D.B., Herschman, H.R., Gilliland, D.G., and Collier, R.J. Epidermal growth factor — toxin A chain conjugates: EGF-ricin A is a potent toxin while EGF-diphtheria fragment A is nontoxic. Cell, 22:563, 1980.PubMedCrossRefGoogle Scholar
  36. 34.
    Klein, G., Giovanella, B.C., Lindahl, T., Fialkow, P.J., Singh, S., and Stehlin, J.S. Direct evidence for the presence of Epstein-Barr virus DNA and nuclear antigen in malignant epithelial cells from patients with poorly differentiated carcinoma of nasopharynx. Proc. Natl. Acad. Sci. USA, 71:4737, 1974.PubMedCrossRefGoogle Scholar
  37. 35.
    Calnek, B.W. and Hitchner, S.B. Localization of viral antigen in chickens infected with Marek’s disease herpesvirus. J. Natl. Cancer Inst., 43:935, 1969.PubMedGoogle Scholar
  38. 36.
    Nazerran, K. and Witter, R.L. Cell-free transmission and in vivo replication of Marek’s disease virus. J. Virol., 5:388, 1970.Google Scholar
  39. 37.
    Andersson, U., Bird, A.G., Britton, S., and Palacios, R. Humoral and cellular immunity studied at the cell level from birth to two years of age. Immunological Rev., 57:5, 1981.CrossRefGoogle Scholar
  40. 38.
    Miller, G. and Lipman, M. Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc. Natl. Acad. Sci. USA, 70:190, 1973.PubMedCrossRefGoogle Scholar
  41. 39.
    Mark, W.H. Transformation of adult human lymphocytes in vitro by Epstein-Barr virus. Doctoral thesis, University of Wisconsin, Madison, 1981.Google Scholar
  42. 40.
    Zinkernagel, R.M. and Rosenthal, K.L. Experiments and speculation on anti-viral specificity of T and B cells. Immunological Rev., 58:131, 1981.CrossRefGoogle Scholar
  43. 41.
    Henle, W. and Henle, G. Seroepidemiology of the virus. In; M.A. Epstein and B.G. Achong (eds.), The Epstein-Barr Virus, pp. 61–78. Berlin: Springer-Verlag, 1979.CrossRefGoogle Scholar
  44. 42.
    Purtilo, D.T., Sakamoto, K., Saemundsen, A.K., Sullivan, J.L., Synnerholm, A-C., Anvret, M., Pritchard, J., Sloper, C., Seiff, C., Pincott, J., Pachman, L., Rich, K., Cruzi, F., Cornet, J., Collins, R., Barnes, N., Knight, J., Sandstedt, B., and Klein, G. Documentation of Epstein-Barr virus in immunodeficient patients with life-threatening lymphoproliferative diseases by clinical, virological and immunopathological studies. Cancer Res., 41:4226, 1981.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1984

Authors and Affiliations

  • Bill Sugden
    • 1
  1. 1.McArdle LaboratoryUniversity of WisconsinMadisonUSA

Personalised recommendations