Advertisement

Selection of Human T-Cell Hybridomas That Produce Inflammatory Lymphokines by the Emetine-Actinomycin D Method

  • Toshiaki Osawa
  • Yoshiro Kobayashi
  • Makoto Asada
  • Masahiro Higuchi
  • Shu-ichi Tsuchiya

Abstract

T lymphocytes produce various physiologically active lymphokines upon stimulation with antigens or mitogens. These lymphokines can roughly be divided into two groups: cell regulatory lymphokines and inflammatory lymphokines. The lymphokines in the former group play roles in the effector mechanism of the immune response; interleukin 2, B-cell growth factor, and T-cell replacing factors belong to this group. Those in the latter group (Table I) react with macrophages and other inflammatory cells or with the vascular endothelium and are considered to be involved in the induction of delayed-type hypersensitivity. However, since activated lymphocytes produce a mixture of many inflammatory lymphokines in very tiny amounts (possibly ~1–10 ng from 106 activated lymphocytes), it has been very difficult to prove directly their participation in the development of various diseases related to delayed hypersensitivity, although some of these lymphokines have actually been detected in tissues or fluids of patients or lesion-bearing experimental animals (Honda and Hayashi, 1982; Cohen and Yoshida, 1983). Furthermore, this limited availability of inflammatory lymphokines has hampered their biochemical characterization and presented several controversial problems concerning the molecular identity between lymphokines. For example, migration-inhibitory factor (MIF) and macrophage-activating factor (MAF), and MAF and immune interferon (IFN-γ), have been assumed to represent identical molecular species, respectively, and skin-reactive factor has been considered to be not a single entity, but a mixture of various lymphokines, including MIF, macrophage chemotactic factor, and vascular permeability factor.

Keywords

Migration Inhibitory Factor Macrophage Activation Factor Macro Phage Distinct Molecular Species Migration Inhibitory Factor Activity 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Cameron, D. J., and Churchill, W. H., 1980, Cytotoxicity of human macrophages for tumor cells: Enhancement by bacterial lipopolysaccharides (LPS), J. Immunol. 124: 708–712.PubMedGoogle Scholar
  2. Churchill, W. H., Jr., Piessens, W. F., Sulis, C. A., and David, J. R., 1975, Macrophages activated as suspension cultures with lymphocyte mediators devoid of antigen become cytotoxic for tumor cells, J. Immunol. 115: 781–786.PubMedGoogle Scholar
  3. Cohen, S., and Yoshida, T., 1983, Physiological and pathological roles of lymphokines, in: Humoral Factors in Host Defence ( Y. Yamamura, H. Hayashi, T. Honjo, T. Kishimoto, M. Muramatsu, and T. Osawa, eds.), Academic Press, New York, pp. 245–256.Google Scholar
  4. Erickson, K. L., Cicurel, L., Gruys, E., and Fidler, I. J., 1982, Murine T-cell hybridomas that produce lymphokine with macrophage-activating factor activity as a constitutive product, Cell. Immunol. 72: 195–201.PubMedCrossRefGoogle Scholar
  5. Foung, S. K. H., Sasaki, D. T., Grumet, F. C., and Engleman, E. G., 1982, Production of functional human T-T hybridomas in selection medium lacking aminopterin and thymidine, Proc. Natl. Acad. Sci. USA 79: 7484–7488.PubMedCrossRefGoogle Scholar
  6. Fox, R. M., Tripp, E. H., and Tattersall, M. H. N., 1980, Mechanism of deoxycytidine rescue of thymidine toxicity in human T-leukemic lymphocytes, Cancer Res. 40: 1718–1721.PubMedGoogle Scholar
  7. Grollman, A. P., 1968, Inhibitors of protein biosynthesis. V. Effects of emetine on protein and nucleic acid biosynthesis in HeLa cells, J. Biol. Chem. 243: 4089–4094.PubMedGoogle Scholar
  8. Hammerstr¢m, J., 1979, In vitro influence of endotoxin on human mononuclear phagocyte structure and function. 2. Enhancement of the expression of cytostatic and cytolytic activity of normal and lymphokine-activated monocytes, Acta Pathol. Microbiol. Immunol. Scand. C 87: 391–399.Google Scholar
  9. Harrington, J. R., Jr., and Stastny, P., 1973, Macrophage migration from an agarose droplet: Development of a micromethod for assay of delayed hypersensitivity, J. Immunol. 110: 752–759.PubMedGoogle Scholar
  10. Henry, W. M., 1981, Interaction of Leishmania with a macrophage cell line. Correlation between intracellular killing and the generation of oxygen intermediates, J. Exp. Med. 153: 1690–1695.CrossRefGoogle Scholar
  11. Higuchi, M., Asada, M., Kobayashi, Y., and Osawa, T., 1983, Human T cell hybridomas producing migration inhibitory factor and macrophage activating factors, Cell. Immunol. 78: 236–248.CrossRefGoogle Scholar
  12. Higuchi, M., Nakamura, N., Tsuchiya, S., Kobayashi, Y., and Osawa, T., 1984, Macrophage activating factor for cytotoxicity produced by a human T cell hybridoma, Cell. Immunol. 87: 626–636.PubMedCrossRefGoogle Scholar
  13. Honda, M., and Hayashi, H., 1982, Characterization of three macrophage chemotactic factors from PPD-induced delayed hypersensitivity reaction sites in guinea pigs, with special reference to a chemotactic lymphokine, Am. J. Pathol. 108: 171–183.PubMedGoogle Scholar
  14. Irigoyen, O., Rizzolo, P. V., Thomas, Y., Rogozinski, L., and Chess, L., 1981, Generation of functional human T cell hybrids, J. Exp. Med. 154: 1827–1837.PubMedCrossRefGoogle Scholar
  15. Kasahara, T., and Shioiri-Nakano, K., 1976, Splenic suppressing factor: Purification and characterization of a factor suppressing thymidine incorporation into activated lymphocytes, J. Immunol. 116: 1251–1256.PubMedGoogle Scholar
  16. Kniep, E. M., Domzig, W., Lohmann-Matthes, M.-L., and Kickhöfen, B., 1981, Partial purification and chemical characterization of macrophage cytotoxicity factor (MCF, MAF) and its separation from migration inhibitory factor (MIF), J. Immunol. 127: 417–422.PubMedGoogle Scholar
  17. Kobayashi, Y., Asada, M., Higuchi, M., and Osawa, T., 1982, Human T cell hybridomas producing lymphokine. I. Establishment and characterization of human T-cell hybridomas producing lymphotoxin and migration inhibitory factor, J. Immunol. 128: 2714–2718.PubMedGoogle Scholar
  18. Köhler, G., and Milstein, C., 1975, Continuous cultures of fused cells secreting antibody of predefined specificity, Nature 256: 495–497.PubMedCrossRefGoogle Scholar
  19. Le, J., Prensky, W., Yip, Y. K., Chang, Z., Hoffman, T., Stevenson, H. C., Balazs, I., Sadlik, J. R., and Vilcek, J., 1983, Activation of human monocyte cytotoxicity by natural and recombinant immune interferon, J. Immunol. 131: 2821–2826.PubMedGoogle Scholar
  20. Mayer, L., Fu, S. M., and Kunkell, H. G., 1982, Human T cell hybridomas secreting factors for IgAspecific help, polyclonal B cell activation, and B cell proliferation, J. Exp. Med. 156: 1860–1865.PubMedCrossRefGoogle Scholar
  21. Meltzer, M. S., 1981, Tumor cytotoxicity by lymphokine-activated macrophages: Development of macrophage tumoricidal activity requires a sequence of reactions, Lymphokines 3: 319–343.Google Scholar
  22. Nathan, C. F., Murray, H. W., Wiebe, M. E., and Rubin, B. Y., 1983, Identification of interferony as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity, J. Exp. Med. 158: 670–689.CrossRefGoogle Scholar
  23. Onozaki, K., Haga, S., Ichikawa, M., Homma, Y., Miura, K., and Hashimoto, T., 1981, Production of an antibody against guinea pig MIF. III. Biological activity of MIF recovered from immunadsorbent column chromatography, Cell Immunol. 61: 165–175.PubMedCrossRefGoogle Scholar
  24. Pace, J. L., Russell, S. W., Torres, B. A., Johnson, H. M., and Gray, P. W., 1983, Recombinant mouse y interferon induces the priming step in macrophage activation for tumor cell killing, J. Immunol. 130: 2011–2013.PubMedGoogle Scholar
  25. Perry, R. P., 1963, Selective effects of actinomycin D on the intracellular distribution of RNA synthesis in tissue culture cells, Exp. Cell Res. 29: 400–406.CrossRefGoogle Scholar
  26. Ratliff, T. L., Thomasson, D. L., McCool, R. E., and Catalona, W. J., 1982a, Production of macrophage activation factor by a T-cell hybridoma, Cell. Immunol. 68: 311–321.PubMedCrossRefGoogle Scholar
  27. Ratliff, T. L., Thomasson, D. L., McCool, R. E., and Catalona, W. J., 1982b, T-cell hybridoma production of macrophage activation factor (MAF). 1. Separation of MAF from interferon gamma, J. Reticuloendothel. Soc. 31: 393–397.PubMedGoogle Scholar
  28. Schreiber, R. D., Altman, A., and Katz, D. H., 1982, Identification of a T cell hybridoma that produces large quantities of macrophage activating factor, J. Exp. Med. 156: 677–689.PubMedCrossRefGoogle Scholar
  29. Schreiber, R. D., Pace, J. L., Russell, S. W., Altman, A., and Katz, D. H., 1983, Macrophageactivating factor produced by a T cell hybridoma: Physiochemical and biosynthetic resemblance to -y-interferon, J. Immunol. 131: 826–832.PubMedGoogle Scholar
  30. Svedersky, L. P., Benton, C. V., Berger, W. H., Rinderknecht, E., Harkins, R. N., and Palladino, M. A., 1984, Biological and antigenic similarities of murine interferon-y and macrophage-activating factor, J. Exp. Med. 159: 812–827.PubMedCrossRefGoogle Scholar
  31. Weinberg, J. B., Chapman, H. A., Jr., and Hibbs, J. B., Jr., 1978, Characterization of the effects of endotoxin on macrophage tumor cell killing, J. Immunol. 121: 72–80.PubMedGoogle Scholar
  32. Wing, E. J., Waheed, A., Shadduck, K., Nagle, L. S., and Stephenson, K., 1982, Effect of colony stimulating factor on murine macrophages. Induction of antitumor activity, J. Clin. Invest. 69: 270–276.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Toshiaki Osawa
    • 1
  • Yoshiro Kobayashi
    • 1
  • Makoto Asada
    • 1
  • Masahiro Higuchi
    • 1
  • Shu-ichi Tsuchiya
    • 1
  1. 1.Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical SciencesUniversity of TokyoTokyoJapan

Personalised recommendations