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Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1988 Jan;71(1):187–192.

Induction of IgM secretion by chronic B-lymphocytic leukaemia cells in serum-free medium: effects of interferon-alpha, -gamma and phorbol ester.

T H Tötterman 1, M Carlsson 1, K Nilsson 1
PMCID: PMC1541627  PMID: 2450704

Abstract

Serum-free culture conditions would be preferable when studying the cellular and molecular regulation of B lymphocyte activation, proliferation and differentiation. We describe here the morphological and functional differentiation of chronic B-lymphocytic leukaemia (B-CLL) cells from 10 patients cultured in serum-free medium. When exposed to the phorbol ester TPA, cells from 8/10 cases expressed blastoid morphology and secreted significant levels of monoclonal IgM. The addition of 0.5% newborn calf serum to the serum-free medium increased both the spontaneous and TPA-induced IgM secretion of B-CLL cells by a factor of 6 and 7, respectively. Compared with TPA, significant but lower levels of IgM secretion and morphological differentiation were observed with native purified leucocyte interferon-alpha (IFN-alpha) (6/8 patients), some batches of recombinant IFN-alpha 2 (5/8 patients) and recombinant IFN-gamma (4/8 patients) in a dose-dependent and specific manner. Preactivation of B-CLL cells with TPA or anti-mu antibody was not necessary for the IFN-induced functional maturation. Significant DNA synthesis was not observed with any of the inducers used. These studies show that B-CLL cells can be induced to differentiate under serum-free conditions in response to physiological and non-physiological ligands.

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Selected References

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

  1. Danersund A., Tötterman T. H., Nilsson K., Egle-Jansson I., Kabelitz D., Sjöberg O. Phorbol ester-induced differentiation of chronic B lymphocytic leukaemia cells--regulatory impact of autologous and allogeneic accessory cells. Clin Exp Immunol. 1985 Mar;59(3):644–652. [PMC free article] [PubMed] [Google Scholar]
  2. Farrant J., Newton C. A., North M. E., Weyman C., Brenner M. K. Production of antibody by human B cells under serum-free conditions. J Immunol Methods. 1984 Mar 30;68(1-2):25–34. doi: 10.1016/0022-1759(84)90133-9. [DOI] [PubMed] [Google Scholar]
  3. Farrar W. L., Johnson H. M., Farrar J. J. Regulation of the production of immune interferon and cytotoxic T lymphocytes by interleukin 2. J Immunol. 1981 Mar;126(3):1120–1125. [PubMed] [Google Scholar]
  4. Forsbeck K., Hellman L., Danersund A., Totterman T. H., Pettersson U., Nilsson K. TPA-induced differentiation of chronic lymphocytic leukemia cells: studies on mu-chain expression. Leukemia. 1987 Jan;1(1):38–43. [PubMed] [Google Scholar]
  5. Gordon J., Rowe M., Walker L., Guy G. Ligation of the CD23,p45 (BLAST-2,EBVCS) antigen triggers the cell-cycle progression of activated B lymphocytes. Eur J Immunol. 1986 Sep;16(9):1075–1080. doi: 10.1002/eji.1830160908. [DOI] [PubMed] [Google Scholar]
  6. Härfast B., Huddlestone J. R., Casali P., Merigan T. C., Oldstone M. B. Interferon acts directly on human B lymphocytes to modulate immunoglobulin synthesis. J Immunol. 1981 Nov;127(5):2146–2150. [PubMed] [Google Scholar]
  7. Iscove N. N., Melchers F. Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes. J Exp Med. 1978 Mar 1;147(3):923–933. doi: 10.1084/jem.147.3.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson H. M., Torres B. A. Phorbol ester replacement of helper cell and interleukin 2 requirements in gamma interferon production. Infect Immun. 1982 Jun;36(3):911–914. doi: 10.1128/iai.36.3.911-914.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kehrl J. H., Muraguchi A., Butler J. L., Falkoff R. J., Fauci A. S. Human B cell activation, proliferation and differentiation. Immunol Rev. 1984 Apr;78:75–96. doi: 10.1111/j.1600-065x.1984.tb00477.x. [DOI] [PubMed] [Google Scholar]
  10. Kishimoto T. Factors affecting B-cell growth and differentiation. Annu Rev Immunol. 1985;3:133–157. doi: 10.1146/annurev.iy.03.040185.001025. [DOI] [PubMed] [Google Scholar]
  11. Leibson H. J., Gefter M., Zlotnik A., Marrack P., Kappler J. W. Role of gamma-interferon in antibody-producing responses. 1984 Jun 28-Jul 4Nature. 309(5971):799–801. doi: 10.1038/309799a0. [DOI] [PubMed] [Google Scholar]
  12. Melchers F., Andersson J. Factors controlling the B-cell cycle. Annu Rev Immunol. 1986;4:13–36. doi: 10.1146/annurev.iy.04.040186.000305. [DOI] [PubMed] [Google Scholar]
  13. Nakagawa T., Hirano T., Nakagawa N., Yoshizaki K., Kishimoto T. Effect of recombinant IL 2 and gamma-IFN on proliferation and differentiation of human B cells. J Immunol. 1985 Feb;134(2):959–966. [PubMed] [Google Scholar]
  14. Nakagawa T., Nakagawa N., Volkman D. J., Fauci A. S. Sequential synergistic effect of interleukin 2 and interferon-gamma on the differentiation of a Tac-antigen-positive B cell line. J Immunol. 1986 Jan;136(1):164–168. [PubMed] [Google Scholar]
  15. Neubauer R. H., Goldstein L., Rabin H., Stebbing N. Stimulation of in vitro immunoglobulin production by interferon-alpha. J Immunol. 1985 Jan;134(1):299–304. [PubMed] [Google Scholar]
  16. Ostlund L., Einhorn S., Robèrt K. H., Juliusson G., Biberfeld P. Chronic B-lymphocytic leukemia cells proliferate and differentiate following exposure to interferon in vitro. Blood. 1986 Jan;67(1):152–159. [PubMed] [Google Scholar]
  17. Rodriguez M. A., Prinz W. A., Sibbitt W. L., Bankhurst A. D., Williams R. C., Jr alpha-Interferon increases immunoglobulin production in cultured human mononuclear leukocytes. J Immunol. 1983 Mar;130(3):1215–1219. [PubMed] [Google Scholar]
  18. Romagnani S., Giudizi G. M., Almerigogna F., Biagiotti R., Alessi A., Mingari C., Liang C. M., Moretta L., Ricci M. Analysis of the role of interferon-gamma, interleukin 2 and a third factor distinct from interferon-gamma and interleukin 2 in human B cell proliferation. Evidence that they act at different times after B cell activation. Eur J Immunol. 1986 Jun;16(6):623–629. doi: 10.1002/eji.1830160607. [DOI] [PubMed] [Google Scholar]
  19. Rowe M., Hildreth J. E., Rickinson A. B., 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. 1982 Apr 15;29(4):373–381. doi: 10.1002/ijc.2910290403. [DOI] [PubMed] [Google Scholar]
  20. Sidman C. L., Marshall J. D., Shultz L. D., Gray P. W., Johnson H. M. Gamma-interferon is one of several direct B cell-maturing lymphokines. 1984 Jun 28-Jul 4Nature. 309(5971):801–804. doi: 10.1038/309801a0. [DOI] [PubMed] [Google Scholar]
  21. Smeland E., Funderud S., Ruud E., Kiil Blomhoff H., Godal T. Characterization of two murine monoclonal antibodies reactive with human B cells. Their use in a high-yield, high-purity method for isolation of B cells and utilization of such cells in an assay for B-cell stimulating factor. Scand J Immunol. 1985 Mar;21(3):205–214. doi: 10.1111/j.1365-3083.1985.tb01422.x. [DOI] [PubMed] [Google Scholar]
  22. Suzuki T., Butler J. L., Cooper M. D. Human B cell responsiveness to B cell growth factor after activation by phorbol ester and monoclonal anti-mu antibody. J Immunol. 1985 Apr;134(4):2470–2476. [PubMed] [Google Scholar]
  23. Tanno Y., Arai S., Takishima T. Induction of immunoglobulin-producing human peripheral blood lymphocytes in serum-free medium. J Immunol Methods. 1982 Jul 30;52(2):255–265. doi: 10.1016/0022-1759(82)90052-7. [DOI] [PubMed] [Google Scholar]
  24. Touraine J. L., Hadden J. W., Touraine F., Hadden E. M., Estensen R., Good R. A. Phorbol myristate acetate: a mitogen selective for a T-lymphocyte subpopulation. J Exp Med. 1977 Feb 1;145(2):460–465. doi: 10.1084/jem.145.2.460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tötterman T. H., Nilsson K., Sundström C. Phorbol ester-induced differentiation of chronic lymphocytic leukaemia cells. Nature. 1980 Nov 13;288(5787):176–178. doi: 10.1038/288176a0. [DOI] [PubMed] [Google Scholar]

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