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Lymphocytolytic activity of glucocorticoids in vitro:51Cr release under equilibrium conditons

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Summary

Cytolytic activity of glucocorticoids in vitro is assessed by measuring radiochromium release from steroid-treated thymic lymphocytes under the equilibrium conditions provided by a continuous-labeling technique. Isotope release is a glucocorticoid-specific effect produced at physiological concentrations and is virtually abolished by inhibitors of RNA and protein synthesis. The relative lytic potencies of the steroids tested are comparable to those reported for glucocorticoids as measured by other methods. This procedure not only possesses the advantages typical of isotopic techniques in general, but, in addition, circumvents the problem of “spontaneous” label release associated with the pulse-labeling method. It is a useful alternative to the morphologic examination of cells or the estimation of cell viability for determination of glucocorticoid cytolytic activity.

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References

  1. Munck, A., and K. Leung. 1977. Glucocorticoid receptors and mechanisms of action. In: J. R. Pasqualini (Ed.),Receptors and Mechanism of Action of Steroid Hormones. Marcel Dekker Inc., New York, pp. 311–397.

    Google Scholar 

  2. Munck, A., and D. A. Young. 1975. Corticosteroids and lymphoid tissue. In: H. Blaschko, G. Sayers, and A. D. Smith (Eds.),Handbook of Physiology. Vol. VI. American Physiological Society, Washington, pp. 231–243.

    Google Scholar 

  3. Burton, A. F., J. M. Storr, and W. L. Dunn. 1967. Cytolytic action of corticosteroids on thymus and lymphoma cellsin vitro. Can. J. Biochem. 45: 289–297.

    PubMed  CAS  Google Scholar 

  4. Claman, H. N., J. W. Moorhead, and W. H. Benner. 1971. Corticosteroids and lymphoid cellsin vitro. I. Hydrocortisone lysis of human, guinea pig, and mouse thymus cells. J. Lab. Clin. Med. 78: 499–507.

    PubMed  CAS  Google Scholar 

  5. Giddings, S. J., and D. A. Young. 1974. Anin vitro effect of physiological levels of cortisol and related steroids on the structural integrity of the nucleus in rat thymic lymphocytes as measured by resistance to lysis. J. Steroid Biochem. 5: 587–595.

    Article  PubMed  CAS  Google Scholar 

  6. Schrek, R. 1949. Cytotoxic action of hormones of the adrenal cortex according to the method of unstained cell counts. Endocrinology 45: 317–334.

    CAS  PubMed  Google Scholar 

  7. Leung, K., and A. Munck. 1975. Long-term incubation of rat thymus cells: Cytolytic actions of glucocorticoidsin vitro. Endocrinology 97: 744–748.

    PubMed  CAS  Google Scholar 

  8. Stuart, J. J., and M. Ingram. 1971. The effect of cortisol on viability and glucose uptake in rat thymocytesin vitro. Proc. Soc. Exp. Biol. Med. 136: 1146–1150.

    PubMed  CAS  Google Scholar 

  9. Phillips, H. J. 1973. Dye exclusion tests for cell viability. In: P. F. Kruse, Jr., and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 406–408.

    Google Scholar 

  10. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265–275.

    PubMed  CAS  Google Scholar 

  11. Shortman, K., N. Williams, and P. Adams. 1972. The separation of different cell classes from lymphoid organs. V. Simple procedures for the removal of cell debris, damaged cells and erythroid cells from lymphoid cell suspensions. J. Immunol. Methods 1: 273–287.

    Article  PubMed  CAS  Google Scholar 

  12. Makman, M. H., S. Nakagawa, B. Dvorkin, and A. White. 1970. Inhibitory effects of cortisol and antibiotics on substrate entry and ribonucleic acid synthesis in rat thymocytesin vitro. J. Biol. Chem. 245: 2556–2563.

    PubMed  CAS  Google Scholar 

  13. Kleine, N., and H. Heimpel. 1965. The early loss of radioactivity in51Cr survival curves: Destruction of cells or loss of the label? Blood 26: 819–822.

    PubMed  CAS  Google Scholar 

  14. Mollison, P. L. 1961. Further observations on the normal survival curve of51Cr-labelled red cells. Clin. Sci. 21: 21–36.

    PubMed  CAS  Google Scholar 

  15. Hughes Jones, N. C., and P. L. Mollison. 1956. The interpretation of measurements with51Cr-labelled red cells. Clin. Sci. 15: 207–218.

    Google Scholar 

  16. Donohue, M., A. G. Motulsky, E. R. Giblett, G. Pirzio-Biroli, V. Viranuvatti, and C. A. Finch. 1955. The use of chromium as a red-cell tag. Br. J. Haematol. 1: 249–263.

    PubMed  CAS  Google Scholar 

  17. Ebaugh, F. G., Jr., C. P. Emerson, and J. F. Ross. 1953. The use of radioactive chromium 51 as an erythrocyte tagging agent for the determination of red cell survivalin vivo. J. Clin. Invest. 32: 1260–1276.

    PubMed  CAS  Google Scholar 

  18. Strumia, M. M., A. Dugan, and L. S. Colwell. 1962. The measure of erythropoiesis in anemias. II. The immediate and subsequent loss of transfused erythrocytes in healthy subjects. Blood 19: 115–122.

    PubMed  CAS  Google Scholar 

  19. Eadie, G. S., and I. W. Brown, Jr. 1955. The potential life span and ultimate survival of fresh red blood cells in normal healthy recipients as studied by simultaneous Cr51 tagging and differential hemolysis. J. Clin. Invest. 34: 629–636.

    Article  PubMed  CAS  Google Scholar 

  20. McMillan, R., and J. L. Scott. 1968. Leukocyte labeling with51chromium. I. Technic and results in normal subjects. Blood 32: 738–754.

    PubMed  CAS  Google Scholar 

  21. Bainbridge, D. R., L. Brent, and G. Gowland. 1966. Distribution of allogeneic51Cr-labelled lymph node cells in mice. Transplantation 4: 138–153.

    Article  PubMed  CAS  Google Scholar 

  22. Sanderson, A. R. 1964. Cytotoxic reactions of mouse iso-antisera: Preliminary considerations. Br. J. Exp. Pathol. 45: 398–408.

    PubMed  CAS  Google Scholar 

  23. Keller, R. 1973. Cytostatic elimination of syngeneic rat tumor cellsin vitro by nonspecifically activated macrophages. J. Exp. Med. 138: 625–644.

    Article  PubMed  CAS  Google Scholar 

  24. Rajam, P. C., A.-L. Jackson, and S. H. Black. 1958. The intracellular labeling of Ehrlich mouse ascites tumor cells with radiochromate. J. Lab. Clin. Med. 51: 767–772.

    PubMed  CAS  Google Scholar 

  25. Holm, G., and P. Perlmann. 1967. Quantitative studies on phytohaemagglutinin-induced cytotoxicity by human lymphocytes against homologous cells in tissue culture. Immunology 12: 525–536.

    PubMed  CAS  Google Scholar 

  26. Nakazawa, S., J. Minowada, T. Tsubota, and L. F. Sinks. 1978. Profound radiosensitivity in “leukemic” T-cell lines and T-cell-type acute lymphoblastic leukemia demonstrated by sodium [51Cr]chromate labeling. Cancer Res. 38: 1661–1666.

    PubMed  CAS  Google Scholar 

  27. Barr, R. D., W. Doughtery, and S. Perry. 1976. A reappraisal of cellular labeling with radiochromium salts. J. Lab. Clin. Med. 88: 1032–1040.

    PubMed  CAS  Google Scholar 

  28. Sanderson, C. J. 1976. The uptake and retention of chromium by cells. Transplantation 21: 526–529.

    PubMed  CAS  Google Scholar 

  29. Ronai, P. M. 1969. The elution of51Cr from labelled leukocytes—a new theory. Blood 33: 408–413.

    PubMed  CAS  Google Scholar 

  30. Rajam, P. C., and A.-L. Jackson 1958. Distribution and valence state of radiochromium in intracellularly labeled Ehrlich mouse ascites carcinoma cells. Proc. Soc. Exp. Biol. Med. 99: 210–213.

    PubMed  CAS  Google Scholar 

  31. Skrabut, E. M., N. Catsimpoolas, J. P. Crowley, and C. R. Valeri. 1976. (51Cr)Sodium chromate incorporation into the soluble fraction of the human erythrocyte: Binding not associated with the hemoglobin monomeric subunit. Biochem. Biophys. Res. Commun. 69: 672–677.

    Article  PubMed  CAS  Google Scholar 

  32. Scaife, J. F., and P. V. Vittorio. 1964. The use of chromium-51 as a sensitive quantitative criterion of early radiation damage to rat thymocytes. Can. J. Biochem. 42: 503–512.

    Article  PubMed  CAS  Google Scholar 

  33. Pearson, H. A. 1963. The binding of Cr51 to hemoglobin. I.In vitro studies. Blood 22: 218–230.

    PubMed  CAS  Google Scholar 

  34. Mollison, P. L., and N. Veall. 1955. The use of the isotope51Cr as a label for red cells. Br. J. Haematol. 1: 62–74.

    PubMed  CAS  Google Scholar 

  35. Zawydiwski, R., and G. R. Duncan. 1978. Spontaneous51Cr release by isolated rat hepatocytes: An indicator of membrane damage. In Vitro 14: 707–714.

    Article  PubMed  CAS  Google Scholar 

  36. Ormos, G., and S. Mányai. 1974. Chromate uptake by human red blood cells: Comparison of permeability for different divalent anions. Acta Biochim. Biophys. 9: 197–207.

    CAS  Google Scholar 

  37. Abrahamsen, A. F. 1968. A modification of the technique for51Cr-labelling of blood platelets giving increased circulating platelet radioactivity. Scand. J. Haematol. 5: 53–63.

    Article  PubMed  CAS  Google Scholar 

  38. Burton, A. F. 1965. Inhibition of 11β-hydroxysteroid dehydrogenase activity in rat and mouse tissuesin vitro andin vivo. Endocrinology 77: 325–331.

    Article  PubMed  CAS  Google Scholar 

  39. Williams, H. E., and C. E. Becher. 1972. Endocrine disorders. In: K. L. Melmon, and H. F. Morrelli (Eds.),Clinical Pharmacology: Basic Principles in Therapeutics. Macmillan Publishing Co., Inc., New York, pp. 290–381.

    Google Scholar 

  40. Russell, J. A., 1965. The adrenals. In: T. C. Ruch, and H. D. Patton (Eds.),Physiology and Biophysics. 19th Edition. W. B. Saunders Co., Philadelphia, pp. 1121–1146.

    Google Scholar 

  41. Parker, R. B., and D. R. Waud. 1971. Pharmacological estimation of drug-receptor dissociation constants. Statistical evaluation. I. Agonists. J. Pharmacol. Exp. Ther. 177: 1–12.

    CAS  Google Scholar 

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  1. Faculty of Pharmacy, University of Toronto, M5S 1A1, Toronto, Ontario, Canada

    R. Zawydiwski & G. R. Duncan

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  1. R. Zawydiwski
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  2. G. R. Duncan
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This work was supported by the Medical Research Council of Canada (MA-2996).

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Zawydiwski, R., Duncan, G.R. Lymphocytolytic activity of glucocorticoids in vitro:51Cr release under equilibrium conditons. In Vitro 15, 603–611 (1979). https://doi.org/10.1007/BF02618261

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