Advertisement

Chemical and Biological Properties of Human Lymphotoxin

  • Bharat B. Aggarwal
  • Barbara Moffat
  • Sang He Lee
  • Richard N. Harkins
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)

Abstract

The cytotoxicity of lymphocytes toward allogeneic target cells in vitro was first shown by Govaerts (1960). This was later confirmed by Rosenau and Moon (1961). It was demonstrated that lymphocytes, when activated with antigen or mitogen, secrete a soluble cytotoxin which was named lymphotoxin (Granger and Kolb, 1968). Since then a number of established human lymphoid cell lines have also been reported to produce a similar cytotoxic molecule (Amino et al., 1974; Granger et al., 1970; Shacks et al., 1973; Papermaster et al., 1976). The activity of lymphotoxin has been tested against a number of cell lines both of animal and human origin. It has been demonstrated by several workers that lymphotoxin is both cytostatic and cytolytic to target cells (Evans and Heinbaugh, 1981; Rosenau, 1981; Sawada et al., 1976). Most of the biological studies with lymphotoxin have been performed with relatively crude preparations. The isolation of lymphotoxin has been a rather difficult task due to its heterogeneous nature and also because of the small amounts produced by normal lymphocytes and various lymphoid cell lines. We have purified human lymphotoxin from several hundred liters of cell conditioned medium derived from the lymphoblastoid cell line RPMI 1788. Using purified material we have also examined the in vitro effects of lymphotoxin on human tumor and normal cells.

Keywords

Human Lung Fibroblast Lymphoid Cell Line Adenoid Tissue Electrophoretic Homogeneity Human Lung Carcinoma A549 Cell 
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. Aggarwal, B. B., Moffat, B., and Harkins, R. N., 1982, Purification and characterization of lymphotoxin from human lymphoblastoid cell line 1788, in: Interleukins, Lymphokines and Cytokines (J.J. Oppenheim and S. Cohen, eds.), pp. 521–526, Academic Press, New York.Google Scholar
  2. Amino, N., Linn, E. S., Pysher, T. J., Mier, R., Moore, G. E., and DeGroot, L. J., 1974, Human lymphotoxin obtained from established lymphoid lines: Purification characteristics and inhibition by antiimmunoglobulins, J. Immunol. 113:1334–1345.PubMedGoogle Scholar
  3. Evans, C. H., 1982, Lymphotoxin—An immunologic hormone with anticarcinogenic and antitumor activity, Cancer Immunol. Immunother. 12:181–190.CrossRefGoogle Scholar
  4. Evans, C. H., and Heinbaugh, J. A., 1981, Lymphotoxin cytotoxicity, a combination of cytolytic and cytostatic cellular responses, Immunopharmacology 3:347–359.PubMedCrossRefGoogle Scholar
  5. Gately, M. K., and Mayer, M. M., 1972, The effect of antibodies to complement components C2, C3 and C5 on the production and action of lymphotoxin, J. Immunol. 109:728.PubMedGoogle Scholar
  6. Govaerts, A., 1960, Cellular antibodies in kidney hemotransplantation, J. Immunol. 85:516–522.PubMedGoogle Scholar
  7. Granger, G. A., and Kolb, W. P., 1968, Lymphocyte in vitro cytotoxicity: Mechanism of immune and non-immune small lymphocyte mediated target L-cell destruction, J. Immunol. 101:111–120.PubMedGoogle Scholar
  8. Granger, G. A., and Williams, T. W., 1968, Lymphocyte cytotoxicity in vitro: Activation and release of a cytotoxic factor, Nature (London) 218:1253–1254.CrossRefGoogle Scholar
  9. Granger, G. A., Moore, G. E., White, J. G., Matzinger, P., Sundsmo, J. S., Shupe, S., Kolb, W. P., Kramer, J., and Glade, P. R., 1970, Production of lymphotoxin and migration inhibiting factor by established human lymphocytic cell lines, J. Immunol. 104:1476–1485.PubMedGoogle Scholar
  10. Granger, G. A., Laserna, E. C., Kolb, W. P., and Chapman, F., 1973, Human lymphotoxin: Purification and some properties, Proc. Natl. Acad. Sci. USA 70:27–30.PubMedCrossRefGoogle Scholar
  11. Granger, G. A., Yamamoto, R. S., Fair, D. S., and Hiserodt, J. C., 1978, The human LT system. I. Physical-chemical heterogeneity of LT molecules released by mitogen activated human lymphocytes in vitro, Cell. Immunol. 38:388–402.PubMedCrossRefGoogle Scholar
  12. Khan, A., Weldon, D., Duvall, J., Pichangkul, S., Hill, N. O., Mutx, D., Lanius, R., and Ground, M., 1982a, A standardized automated computer assisted micro-assay for lymphotoxin, in: Human Lymphokines: The Biological Immune Response Modifiers (A. Khan and N. O. Hill, eds.), pp. 23–32, Academic Press, New York.CrossRefGoogle Scholar
  13. Khan, A., Hill, N. O., Ridgway, H., and Webb, K., 1982b, Preclinical and phase I clinical trials with lymphotoxin, in: Human Lymphokines (A. Khan and N. O. Hill, eds.), pp. 621–632, Academic Press, New York.CrossRefGoogle Scholar
  14. Klostergaard, J., and Granger, G. A., 1981, Human lymphotoxin: Purification to electrophoretic homogeneity of the ah receptor bearing class, Mol. Immunol. 18:455–458.PubMedCrossRefGoogle Scholar
  15. Klostergaard, J., Yamamoto, R. S., and Granger, G. A., 1980, Human and murine lymphotoxins as a multi-component system: Progress in the purification of the human al. component, Mol. Immunol. 17:613–623.PubMedCrossRefGoogle Scholar
  16. Klostergaard, J., Long, S., and Granger, G. A., 1981, Purification of human alpha light class lymphotoxin to electrophoretic homogeneity, Mol. Immunol. 18:1049–1054.PubMedCrossRefGoogle Scholar
  17. Kolb, W. P., and Granger, G. A., 1968, Lymphocyte in vitro cytotoxicity: Characterization of human lymphotoxin, Proc. Natl. Acad. Sci. USA 61:1250–1255.PubMedCrossRefGoogle Scholar
  18. Kolb, W. P., and Granger, G. A., 1970, Lymphocyte in vitro cytotoxicity: Characterization of mouse lymphotoxin, Cell. Immunol. 1:122–132.PubMedCrossRefGoogle Scholar
  19. Kramer, J. J., and Granger, G. A., 1972, The in vitro induction and release of cell toxins by immune C57 black 6 mouse peritoneal macrophages, Cell. Immunol. 3:88–100.PubMedCrossRefGoogle Scholar
  20. Kramer, S. L., and Granger, G. A., 1976, The role of lymphotoxin in target cell destruction induced by mitogen activated human lymphocytes in vitro. II. The correlation of temperature and trypsin sensitive phases of lymphotoxin induced and lymphocyte mediated cytotoxicity, J. Immunol. 116:562–567.PubMedGoogle Scholar
  21. Laemmli, U. K., 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T-4, Nature (London) 227:680–685.CrossRefGoogle Scholar
  22. Papermaster, B.W., Holterman, O. A., Klein, E., Parnrett, S., Dobkin, D., Laudico, R., and Djerass, I., 1976, Lymphokine properties of a lymphoid cultured cell supernatant fraction active in promoting tumor regression, Clin. Immunol. Immunopathol 5:48–59.PubMedCrossRefGoogle Scholar
  23. Papermaster, B. W., Smith, M. E., and McEntire, J. E., 1981, Lymphotoxins: Soluble cytotoxic molecules secreted by lymphocytes, in: The Lymphokines: Biochemistry and Biological Activity (J. W. Hadden and W. E. Stewart II, eds.), pp. 149–180, Humana Press, Clifton, N.J.Google Scholar
  24. Peters, J. B., Stratton, J. A., Stempel, K. E., Yu, D., and Cardin, C., 1973, Characteristics of a cytotoxin (“lymphotoxin”) produced by stimulation of human lymphoid tissue, J. Immunol. 111:770–782.Google Scholar
  25. Rosenau, W., 1981, Lymphotoxin: Properties, role, and mode of action, Int. J. Immunopharmacol. 3:1–8.PubMedCrossRefGoogle Scholar
  26. Rosenau, W., and Moon, H. D., 1961, Lysis of homologous cells by sensitized lymphocytes in tissue culture, J. Natl. Cancer Inst. 27:471–483.PubMedGoogle Scholar
  27. Rosenau, W., and Tsoukas, C. D., 1976, Lymphotoxin, A review and analysis, Am. J. Pathol. 84:580–596.PubMedGoogle Scholar
  28. Russell, S. W., Rosenau, W., Goldberg, M. L., and Kumitomi, G., 1972, Purification of human lymphotoxin, J. Immunol. 109:784–790.PubMedGoogle Scholar
  29. Sawada, J., Shioiri-Nakano, K., and Osawa, T., 1976, Cytotoxic activity of purified lymphotoxin against various cell lines, Jpn. J. Exp. Med. 46:263–267.PubMedGoogle Scholar
  30. Shacks, S. J., Chiller, J., and Granger, G. A., 1973, Studies on in vitro models of cellular immunity: The role of T and B cells in the secretion of lymphotoxin, Cell. Immunol. 7:313–321.PubMedCrossRefGoogle Scholar
  31. Smith, M. E., Landico, R., and Papermaster, B.W., 1977, A rapid quantitative assay for lymphotoxin, J. Immunol. Methods 14:243–251.PubMedCrossRefGoogle Scholar
  32. Spofford, B. T., Daynes, R. A., and Granger, G. A., 1974, Cell mediated Immunity in vitro: A highly sensitive assay for human lymphotoxin, J. Immunol. 112:2111–2116.PubMedGoogle Scholar
  33. Walker, S. M., and Lucas, Z. J., 1972, Cytotoxic activity of lymphocytes. I. Assay for cytotoxicity by rubidium exchange at isotopic equilibrium, J. Immunol. 109:1223–1244.PubMedGoogle Scholar
  34. Walker, S. M., Lee, S. C., and Lucas, Z. J., 1976, Cytotoxic activity of lymphocytes. IV. Heterogeneity of cytotoxins in supernatants of mitogen activated lymphocytes, J. Immunol. 116:807–815.PubMedGoogle Scholar
  35. Williams, T. W., and Bellanti, J. A., 1983, In vitro synergism between interferons and human lym-photoxin: Enhancement of lymphotoxin induced target cell killing, J. Immunol. 130:518–520.PubMedGoogle Scholar
  36. Wright, S. C., and Bonavida, B., 1982, Studies on the mechanism of natural killer (NK) cell mediated cytotoxicity (CMC). I. Release of cytotoxic factors specific for NK sensitive target cells (NKCF) during coculture of NK effector cells with NK target cells, J. Immunol. 129:433–439.PubMedGoogle Scholar
  37. Yamamoto, R. S., Hiserodt, J. C., Lewis, J. E., Carmack, C. E., and Granger, G. A., 1978, The human LT system. II. Immunological relationship of LT molecules released by mitogen activated human lymphocytes in vitro. Cell. Immunol. 38:403–416.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Bharat B. Aggarwal
    • 1
  • Barbara Moffat
    • 1
  • Sang He Lee
    • 2
  • Richard N. Harkins
    • 1
  1. 1.Department of Protein BiochemistryGenentech Inc.South San FranciscoUSA
  2. 2.Department of Pharmacological SciencesGenentech Inc.South San FranciscoUSA

Personalised recommendations