Medical Microbiology and Immunology

, Volume 172, Issue 2, pp 87–99 | Cite as

Establishment of spontaneously outgrowing lymphoblastoid cell lines with Cyclosporin A

  • M. v. Knebel Doeberitz
  • G. W. Bornkamm
  • H. zur Hausen


To establish spontaneously outgrowing EBV (Epstein-Barr virus) transformed lymphoid cell lines from healthy EBV-seropositive adults different culture conditions were investigated. The effect of T-cell depletion was compared with that of adding Cyclosporin A (CSA) to unseparated mononuclear blood cells, either at the onset of cultivation or after 3 weeks. The highest frequency of spontaneous outgrowth of EBV-transformed cells was obtained in unseparated cultures when the addition of CSA was delayed. The presence of unimpaired T cells at the onset of the culture period appeared to have a favourable effect on the spontaneous outgrowth of EBV-carrying lymphoblastoid cell lines.


Blood Cell Culture Condition Cyclosporin Culture Period Favourable Effect 
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. Berger R, Meingassner JG, Knapp W (1983) In vitro effects of Cyclosporin A on human B cell responses. Scand J Immunol (in press)Google Scholar
  2. Bird AG, McLachlan SM, Britton S (1981) Cyclosporin A promotes spontaneous outgrowth in vitro of EBV induced B cell lines. Nature 289:300–301CrossRefPubMedGoogle Scholar
  3. Böyum A (1968) Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Lab Invest 21:77PubMedGoogle Scholar
  4. Bunjes D, Hardt C, Röllinghoff M, Wagner H (1981) Cyclosporin A mediates immunosuppression of primary cytotoxic T cell responses by impairing the release of interleukin 1 and 2. Eur J Immunol 11:657–661PubMedGoogle Scholar
  5. Crawford DH, Edwards JMB, Sweny P, Janossy G, Hoffbrand AV (1981) Long-term T cell immunity to EBV in renal-allograft recipients receiving Cyclosporin A. Lancet I:10–12CrossRefGoogle Scholar
  6. Ford RJ, Metha SR, Franzini D, Montagna R, Lachman LB, Maizel AL (1981) Soluble factor activation of human B lymphocytes. Nature 294:261–263CrossRefPubMedGoogle Scholar
  7. Henle G, Henle W (1966) Immunofluorescence in cells derived from Burkitt's lymphoma. J Bacteriol 91:1248–1256PubMedGoogle Scholar
  8. Henle W, Diehl V, Kohn G, zur Hausen H, Henle G (1967) Herpes type virus and chromosome marker in normal leukocytes after growth with irradiated Burkitt cells. Science 157:1064–1065PubMedGoogle Scholar
  9. Henle W, Henle G, Zajac B, Pearson G, Waubke R, Scriba M (1970) Differential reactivity of human sera with EBV-induced “early antigens”. Science 169:188–190PubMedGoogle Scholar
  10. Hess AD, Tutschka J, Santos GW (1982) Effect of Cyclosporin A on human lymphocyte responses in vitro. III. CSA inhibits the production of T lymphocyte growth factors in secondary mixed lymphocyte responses but does not inhibit the response of primed lymphocytes to TCGF. J Immunol 128:355–359PubMedGoogle Scholar
  11. Hinuma Y, Konn J, Yamaguchi DJ, Wudarski JR, Blakeslee J, Grace J (1967) Immunofluorescene and herpes type virus particles in the P3HR-1 Burkitt lymphoma cell line. J Virol 1:1045–1051PubMedGoogle Scholar
  12. Howard MS, Kessler T, Chused T, Paul WE (1981) Long term culture of normal mouse B lymphocytes. Proc Natl Acad Sci USA 78:5788–5792PubMedGoogle Scholar
  13. Kamoun M, Martin PJ, Hansen JA, Brown MA, Siadak AW, Nowinski RC (1981) Identification of a human T-lymphocyte surface protein associated with the E-rosette receptor. J Exp Med 153:207–212CrossRefPubMedGoogle Scholar
  14. Kaplan ME, Clark C (1974) An improved rosetting assay for detection of human T-lymphocytes. J Immunol Methods 5:131–135CrossRefPubMedGoogle Scholar
  15. Klein E, Ernberg I, Masucci MG, Szigeti R, Wu YT, Masucci G, Svedmyr E (1981) T cell responses to B cells and EBV antigens in infectious mononucleosis. Cancer Res 41:4210–4215PubMedGoogle Scholar
  16. Lernhardt W, Corbel C, Wall R, Melchers F (1982) T cell hybridoma which produce B lymphocyte replacing factor only. Nature 300:355–357CrossRefPubMedGoogle Scholar
  17. Lipinski M, Fridman WH, Tursz T, Vincent C, Pilous D, Fellous M (1979) Absence of allogenic restriction in human T cell mediated cytotoxicity to EBV infected target cells. J Exp Med 150:1310–1322CrossRefPubMedGoogle Scholar
  18. Miller G, Lipman M (1973) Release of infectious EBV by transformed marmoset leukocytes. Proc Natl Acad Sci USA 70:190–194PubMedGoogle Scholar
  19. Misko IS, Moss DJ, Pope JH (1980) HLA-antigen related restriction of T lymphocyte cytotoxicity to EBV virus. Proc Natl Acad Sci USA 77:4247–4250PubMedGoogle Scholar
  20. Moss DJ, Rickinson AB, Pope JH (1978) Long term T cell mediated immunity to EBV in man. I. Complete regression of virus induced transformation in cultures of seropositive donor leukocytes. Int J Cancer 22:662–668PubMedGoogle Scholar
  21. Moss DJ, Rickinson AB, Pope JH (1979) Long term T cell mediated immunity to EBV in man. III. Activation of cytotoxic T cells in virus infected leukocyte cultures. Int J Cancer 23:618–625PubMedGoogle Scholar
  22. Moss DJ, Rickinson AB, Wallace LE, Epstein MA (1981) Sequential appearance of EBV nuclear and lymphocyte-detected membrane antigens in B cell transformation Nature 291:664–666CrossRefPubMedGoogle Scholar
  23. Nilsson K, Klein G, Henle G, Henle W (1971) The establishment of lymphoblastoid lines from adult and fetal human lymphoid tissue and its dependence on EBV. Int J Cancer 8:443–450PubMedGoogle Scholar
  24. Palacois R (1981) Cyclosporin A abrogates proliferation of T cells and generation of suppressor and cytotoxic T cell function induced by EBV. Immunobiology 160: 321–329PubMedGoogle Scholar
  25. Palacois R, Möller G (1981) Cyclosporin A blocks receptors for HLA-DR antigen on T cells. Nature 290:792–794CrossRefPubMedGoogle Scholar
  26. Pope JH, Horne MK, Scott W (1968) Transformation of fetal human leukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int J Cancer 3:857–866PubMedGoogle Scholar
  27. Purtilo DT (1980) EBV induced oncogenesis in immune deficient individuals. Lancet I:300–303CrossRefGoogle Scholar
  28. Reedman B, Klein G (1973) Cellular localisation of an EBV associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer 11:499–520PubMedGoogle Scholar
  29. Rickinson AB, Jarvis JE, Crawford DH, Epstein MA (1974) Observations on the type of infection by EBV in peripheral lymphoid cells of patients with infectious mononucleosis. Int J Cancer 14:704–715PubMedGoogle Scholar
  30. Rickinson AB, Epstein MA, Crawford DH (1975) Absence of infectious EBV in blood in acute infectious mononucleosis. Nature 258:236–238CrossRefPubMedGoogle Scholar
  31. Rickinson AB, Finerty S, Epstein MA (1977) Mechanism of the establishment of EBV genome-containing lymphoid cell lines from infectious mononucleosis patients: studies with phosphonoacetate. Int J Cancer 20:861–868PubMedGoogle Scholar
  32. Rickinson AB, Moss DJ, Pope JH (1979) Long-term T cell mediated immunity to EBV in man. II. Components necessary for regression in virus infected leukocyte culture. Int J Cancer 23:610–617PubMedGoogle Scholar
  33. Rickinson AB, Wallace LE, Epstein MA (1980) HLA-restricted T cell recognition of EBV infected B cells. Nature 283:865–867CrossRefPubMedGoogle Scholar
  34. Sugamura K, Tanaka Y, Hinuma Y, (1981) Establishment of cytotoxic T cell lines for EBV-transformed autologous cells. Int J Cancer 28:137–142PubMedGoogle Scholar
  35. Tanaka Y, Sugamura K, Hinuma Y, Sato H, Okochi K (1980) Memory of EBV specific cytotoxic T cells in normal seropositive adults as revealed by an in vitro restimulation method. J Immunol 125:1426–1431PubMedGoogle Scholar
  36. Thorley-Lawson DA (1980) The suppression of EBV virus infection in vitro occurs after infection but before transformation of the cell. J Immunol 124:745–751PubMedGoogle Scholar
  37. Thorley-Lawson DA, Chess L, Strominger JL (1977) Suppression of in vitro EBV infection. J Exp Med 146:495–508CrossRefPubMedGoogle Scholar
  38. Tosato G, Magrath IT, Blaese RM (1982) T cell mediated immunoregulation of EBV-seropositive and EBV-seronegative individuals. J Immunol 128:575–579PubMedGoogle Scholar
  39. Wetzel GD, Swain SL, Dutton RW (1982) A monoclonal T cell replacing activity can act directly on B cells to enhance clonal expansion. J Exp Med 156:306–311CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • M. v. Knebel Doeberitz
    • 1
  • G. W. Bornkamm
    • 1
  • H. zur Hausen
    • 1
  1. 1.Institut für VirologieZentrum für Hygiene der Universität FreiburgFreiburgFederal Republic of Germany

Personalised recommendations