Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1986 Jun;58(2):197–202.

Hydroxyl radical scavengers inhibit human lectin-dependent cellular cytotoxicity.

M Melinn, H McLaughlin
PMCID: PMC1452669  PMID: 3011654

Abstract

The role of oxygen-derived free radicals (ODFR) in lectin-dependent cellular cytotoxicity (LDCC) in humans was investigated. The hydroxyl radical traps thiourea, methanol, ethanol and phenol were effective in inhibiting LDCC, as was DABCO, a singlet oxygen quencher. The proposed pathway of hydroxyl radical production in living cells is either an iron catalysed Haber-Weiss reaction or a Fenton reaction. The effect of inhibitors of these pathways was investigated. The superoxide anion scavengers superoxide dismutase, ferricytochrome c and Tiron were without effect. It was shown that Tiron inhibits the lucigenin-amplified chemiluminescence produced by the action of xanthine oxidase, and also the lucigenin-amplified chemiluminescence produced by activated PMN, suggesting that this agent (Tiron) scavenges intracellular superoxide anion. Catalase gave slight inhibition of LDCC only. The ferric iron chelator desferrioxamine gave no protection of the target cells, while the ferrous chelator, 1,10-phenanthroline, inhibited LDCC and partially prevented the detection of hydroxyl radicals generated by the Fe2+-H2O2 system. Cibacron blue, an agent that inhibits NAD(P)H linked enzymes, also inhibited LDCC. The cyclo-oxygenase inhibitors indomethacin and salicylate were without effect, while the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA) inhibited cytolysis. None of the LDCC inhibitors was cytotoxic to the effector cells or to the target cells, neither did they inhibit lymphocyte-target binding. The findings would suggest that hydroxyl radicals have a role to play in human T-cell mediated cytolysis, either as the active lytic agent or as an epiphenomenon.

Full text

PDF
197

Selected References

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

  1. Bradley T. P., Bonavida B. Mechanism of cell-mediated cytotoxicity at the single cell level. III. Evidence that cytotoxic T lymphocytes lyse both antigen-specific and -nonspecific targets pretreated with lectins or periodate. J Immunol. 1981 Jan;126(1):208–213. [PubMed] [Google Scholar]
  2. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  3. Cleland L. G., Betts W. H., Vernon-Roberts B., Bielicki J. Role of iron and influence of antiinflammatory drugs on oxygen-derived free radical production and reactivity. J Rheumatol. 1982 Nov-Dec;9(6):885–892. [PubMed] [Google Scholar]
  4. Devlin R. G., Lin C. S., Perper R. J., Dougherty H. Evaluation of free radical scavengers in studies of lymphocyte-mediated cytolysis. Immunopharmacology. 1981 Jun;3(2):147–159. doi: 10.1016/0162-3109(81)90016-3. [DOI] [PubMed] [Google Scholar]
  5. Duwe A. K., Werkmeister J., Roder J. C., Lauzon R., Payne U. Natural killer cell-mediated lysis involves an hydroxyl radical-dependent step. J Immunol. 1985 Apr;134(4):2637–2644. [PubMed] [Google Scholar]
  6. Kochman S., Bernard J., Schvartz H., Cazabat A., Thiernesse N., Lavaud F., Caulet T. Failure to distinguish ultrastructurally between T4+ (helper) and T8+ (suppressor/cytotoxic) T-cell subsets. Scand J Immunol. 1984 Feb;19(2):165–173. doi: 10.1111/j.1365-3083.1984.tb00913.x. [DOI] [PubMed] [Google Scholar]
  7. Lewis G. P. Immunoregulatory activity of metabolites of arachidonic acid and their role in inflammation. Br Med Bull. 1983 Jul;39(3):243–248. doi: 10.1093/oxfordjournals.bmb.a071827. [DOI] [PubMed] [Google Scholar]
  8. Mazumder A., Grimm E. A., Rosenberg S. A. Characterization of the lysis of fresh human solid tumors by autologous lymphocytes activated in vitro with phytohemagglutinin. J Immunol. 1983 Feb;130(2):958–964. [PubMed] [Google Scholar]
  9. Mello Filho A. C., Hoffmann M. E., Meneghini R. Cell killing and DNA damage by hydrogen peroxide are mediated by intracellular iron. Biochem J. 1984 Feb 15;218(1):273–275. doi: 10.1042/bj2180273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Motoi S., Aoike A., Kawai K., Amagai T., Kishida T. Human cytotoxic T lymphocytes (CTL) induced by phytohemagglutinin: conditions for CTL generation and effect of interferon. Jpn J Med Sci Biol. 1982 Oct-Dec;35(5-6):221–230. doi: 10.7883/yoken1952.35.221. [DOI] [PubMed] [Google Scholar]
  11. Nathan C. F., Mercer-Smith J. A., Desantis N. M., Palladino M. A. Role of oxygen in T cell-mediated cytolysis. J Immunol. 1982 Nov;129(5):2164–2171. [PubMed] [Google Scholar]
  12. Plaut M., Marone G., Gillespie E. The role of cyclic AMP in modulating cytotoxic T lymphocytes. II. Sequential changes during culture in responsiveness of cytotoxic lymphocytes to cyclic AMP-active agents. J Immunol. 1983 Dec;131(6):2945–2952. [PubMed] [Google Scholar]
  13. Podack E. R., Konigsberg P. J. Cytolytic T cell granules. Isolation, structural, biochemical, and functional characterization. J Exp Med. 1984 Sep 1;160(3):695–710. doi: 10.1084/jem.160.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ramstedt U., Rossi P., Kullman C., Warren E., Palmblad J., Jondal M. Free oxygen radicals are not detectable by chemiluminescence during human natural killer cell cytotoxicity. Scand J Immunol. 1984 May;19(5):457–464. doi: 10.1111/j.1365-3083.1984.tb00954.x. [DOI] [PubMed] [Google Scholar]
  15. Redelman D., Hudig D. The mechanism of cell-mediated cytotoxicity. III. Protease-specific inhibitors preferentially block later events in cytotoxic T lymphocyte-mediated lysis than do inhibitors of methylation or thiol-reactive agents. Cell Immunol. 1983 Oct 1;81(1):9–21. doi: 10.1016/0008-8749(83)90206-x. [DOI] [PubMed] [Google Scholar]
  16. Richmond R., Halliwell B., Chauhan J., Darbre A. Superoxide-dependent formation of hydroxyl radicals: detection of hydroxyl radicals by the hydroxylation of aromatic compounds. Anal Biochem. 1981 Dec;118(2):328–335. doi: 10.1016/0003-2697(81)90590-x. [DOI] [PubMed] [Google Scholar]
  17. Rowley D. A., Halliwell B. Superoxide-dependent formation of hydroxyl radicals from NADH and NADPH in the presence of iron salts. FEBS Lett. 1982 Jun 1;142(1):39–41. doi: 10.1016/0014-5793(82)80214-7. [DOI] [PubMed] [Google Scholar]
  18. Seaman W. E. Human natural killer cell activity is reversibly inhibited by antagonists of lipoxygenation. J Immunol. 1983 Dec;131(6):2953–2957. [PubMed] [Google Scholar]
  19. Suthanthiran M., Solomon S. D., Williams P. S., Rubin A. L., Novogrodsky A., Stenzel K. H. Hydroxyl radical scavengers inhibit human natural killer cell activity. Nature. 1984 Jan 19;307(5948):276–278. doi: 10.1038/307276a0. [DOI] [PubMed] [Google Scholar]
  20. Thorne K. J., Franks D. Importance of oxidative metabolism in T cell cytotoxicity: a comparison of cloned T cells and spleen cells. Immunology. 1983 Dec;50(4):645–650. [PMC free article] [PubMed] [Google Scholar]
  21. Thorne K. J., Svvennsen R. J., Franks D. Role of hydrogen peroxide in the cytotoxic reaction of T lymphocytes. Clin Exp Immunol. 1980 Feb;39(2):486–495. [PMC free article] [PubMed] [Google Scholar]
  22. Treves A. J., Yagoda D., Haimovitz A., Ramu N., Rachmilewitz D., Fuks Z. The isolation and purification of human peripheral blood monocytes in cell suspension. J Immunol Methods. 1980;39(1-2):71–80. doi: 10.1016/0022-1759(80)90295-1. [DOI] [PubMed] [Google Scholar]
  23. Yagawa K., Itoh T., Tomoda A. Effect of transmethylation-reaction and increased levels of cAMP on superoxide generation of guinea-pig macrophages induced with wheat germ agglutinin and phorbor myristate. FEBS Lett. 1983 Apr 18;154(2):383–386. doi: 10.1016/0014-5793(83)80187-2. [DOI] [PubMed] [Google Scholar]
  24. Yam L. T., Li C. Y., Crosby W. H. Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol. 1971 Mar;55(3):283–290. doi: 10.1093/ajcp/55.3.283. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES