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. 1987 Jul;61(3):277–282.

Antibody-dependent cytotoxicity on chicken red blood cell targets mediated by the U937 cell line.

M B Villiers, R H Ward, P J Lachmann
PMCID: PMC1453394  PMID: 3301633

Abstract

We have characterized a model system for the study of antibody-dependent cytotoxicity (ADCC) mediated by human macrophages and monocytes. The U937 cell line is used as a source of effector cells. We confirmed a previous report (Gidlund et al., 1981) that U937 can be activated using PMA to kill in ADCC, and the characteristics of the observed cytotoxicity are described. Activation of effectors was maximal after 20-hr preincubation in the presence of 10 ng/ml PMA. In these conditions, lysis was approximately 70% in 2 hr at an effector to target ratio of 5:1. Activation correlated with the expression of complement receptors CR1 and CR3. No antibody-independent cytotoxicity was observed. The effects of various inhibitors of oxygen species were investigated: the lysis obtained in the above conditions was inhibited at 50% in the presence of either 450 mM dimethyl sulphoxide or 30,000 U/ml catalase, but superoxide dismutase (up to 10,000 U/ml) or ferricytochrome c (up to 2 mM) had no effect. The same inhibition was observed with 40 mM desferrioxamine or with 1 mM 0-phenanthroline, which are both iron scavengers, or in the presence of 300 microM colchicine or 1.5 microM dihydrocytochalasin B, which are two inhibitors of cytoskeletal functions. An identical effect was obtained in the presence of 1 TIU/ml bovine pancreas trypsin inhibitor, whereas soya bean trypsin inhibitor, which is more specific, had no effect up to 5000 BAEE U/ml. No inhibition was seen with protein synthesis inhibitors as cycloheximide or puromycin at 40 micrograms/ml. The significance of these results is discussed.

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Selected References

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

  1. Adams D. O., Johnson W. J., Marino P. A. Mechanisms of target recognition and destruction in macrophage-mediated tumor cytotoxicity. Fed Proc. 1982 Apr;41(6):2212–2221. [PubMed] [Google Scholar]
  2. Adams D. O., Marino P. A. Evidence for a multistep mechanism of cytolysis by BCG-activated macrophages: the interrelationship between the capacity for cytolysis, target binding, and secretion of cytolytic factor. J Immunol. 1981 Mar;126(3):981–987. [PubMed] [Google Scholar]
  3. Anderson C. L., Abraham G. N. Characterization of the Fc receptor for IgG on a human macrophage cell line, U937. J Immunol. 1980 Dec;125(6):2735–2741. [PubMed] [Google Scholar]
  4. Bowman B. U., Jr, Shoeb H. A. Effect of oxygen radical scavengers on K-cell cytolysis. Infect Immun. 1984 Mar;43(3):942–946. doi: 10.1128/iai.43.3.942-946.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Changelian P. S., Jack R. M., Collins L. A., Fearon D. T. PMA induces the ligand-independent internalization of CR1 on human neutrophils. J Immunol. 1985 Mar;134(3):1851–1858. [PubMed] [Google Scholar]
  6. Fleer A., Roos D., von dem Borne A. E., Engelfriet C. P. Cytotoxic activity of human monocytes towards sensitized red cells is not dependent on the generation of reactive oxygen species. Blood. 1979 Aug;54(2):407–411. [PubMed] [Google Scholar]
  7. Gidlund M., Orn A., Pattengale P. K., Jansson M., Wigzell H., Nilsson K. Natural killer cells kill tumour cells at a given stage of differentiation. Nature. 1981 Aug 27;292(5826):848–850. doi: 10.1038/292848a0. [DOI] [PubMed] [Google Scholar]
  8. Gilbert D., Peulve P., Daveau M., Ripoche J., Fontaine M. Modulation of complement receptors of a human monocyte cell line, U-937, during incubation with phorbol myristate acetate: expression of an iC3b-specific receptor (CR3). Eur J Immunol. 1985 Oct;15(10):986–991. doi: 10.1002/eji.1830151005. [DOI] [PubMed] [Google Scholar]
  9. Hall R. E., Muchmore A. V., Decker J. M., Blaese R. M. Induction of cytotoxic effector activity in the HL-60 promyelocytic cell line by incubation with phorbol myristate acetate: a model system of human spontaneous monocyte-mediated cytotoxicity. Cell Immunol. 1983 Feb 15;76(1):58–68. doi: 10.1016/0008-8749(83)90348-9. [DOI] [PubMed] [Google Scholar]
  10. Halliwell B., Gutteridge J. M. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984 Apr 1;219(1):1–14. doi: 10.1042/bj2190001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoe S., Rowley D. A., Halliwell B. Reactions of ferrioxamine and desferrioxamine with the hydroxyl radical. Chem Biol Interact. 1982 Jul 15;41(1):75–81. doi: 10.1016/0009-2797(82)90018-7. [DOI] [PubMed] [Google Scholar]
  12. Hogg N., Ross G. D., Jones D. B., Slusarenko M., Walport M. J., Lachmann P. J. Identification of an anti-monocyte monoclonal antibody that is specific for membrane complement receptor type one (CR1). Eur J Immunol. 1984 Mar;14(3):236–243. doi: 10.1002/eji.1830140307. [DOI] [PubMed] [Google Scholar]
  13. Johnson W. J., Pizzo S. V., Imber M. J., Adams D. O. Receptors for maleylated proteins regulate secretion of neutral proteases by murine macrophages. Science. 1982 Nov 5;218(4572):574–576. doi: 10.1126/science.6289443. [DOI] [PubMed] [Google Scholar]
  14. Klassen D. K., Conkling P. R., Sagone A. L., Jr Activation of monocyte and granulocyte antibody-dependent cytotoxicity by phorbol myristate acetate. Infect Immun. 1982 Mar;35(3):818–825. doi: 10.1128/iai.35.3.818-825.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koren H. S., Anderson S. J., Larrick J. W. In vitro activation of a human macrophage-like cell line. Nature. 1979 May 24;279(5711):328–331. doi: 10.1038/279328a0. [DOI] [PubMed] [Google Scholar]
  16. Larrick J. W., Fischer D. G., Anderson S. J., Koren H. S. Characterization of a human macrophage-like cell line stimulated in vitro: a model of macrophage functions. J Immunol. 1980 Jul;125(1):6–12. [PubMed] [Google Scholar]
  17. Lavie G., Leib Z., Servadio C. The mechanism of human NK cell-mediated cytotoxicity. Mode of action of surface-associated proteases in the early stages of the lytic reaction. J Immunol. 1985 Aug;135(2):1470–1476. [PubMed] [Google Scholar]
  18. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  19. 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]
  20. Minta J. O., Pambrun L. In vitro induction of cytologic and functional differentiation of the immature human monocytelike cell line U-937 with phorbol myristate acetate. Am J Pathol. 1985 Apr;119(1):111–126. [PMC free article] [PubMed] [Google Scholar]
  21. Nathan C. F., Silverstein S. C., Brukner L. H., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. II. Hydrogen peroxide as a mediator of cytotoxicity. J Exp Med. 1979 Jan 1;149(1):100–113. doi: 10.1084/jem.149.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nathan C., Cohn Z. Role of oxygen-dependent mechanisms in antibody-induced lysis of tumor cells by activated macrophages. J Exp Med. 1980 Jul 1;152(1):198–208. doi: 10.1084/jem.152.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ojo E., Wigzell H. Natural killer cells may be the only cells in normal mouse lymphoid cell populations endowed with cytolytic ability for antibody-coated tumour target cells. Scand J Immunol. 1978 Apr;7(4):297–306. doi: 10.1111/j.1365-3083.1978.tb00457.x. [DOI] [PubMed] [Google Scholar]
  24. Redelman D., Hudig D. The mechanism of cell-mediated cytotoxicity. I. Killing by murine cytotoxic T lymphocytes requires cell surface thiols and activated proteases. J Immunol. 1980 Feb;124(2):870–878. [PubMed] [Google Scholar]
  25. Ross G. D., Medof M. E. Membrane complement receptors specific for bound fragments of C3. Adv Immunol. 1985;37:217–267. doi: 10.1016/s0065-2776(08)60341-7. [DOI] [PubMed] [Google Scholar]
  26. Ross G. D., Thompson R. A., Walport M. J., Springer T. A., Watson J. V., Ward R. H., Lida J., Newman S. L., Harrison R. A., Lachmann P. J. Characterization of patients with an increased susceptibility to bacterial infections and a genetic deficiency of leukocyte membrane complement receptor type 3 and the related membrane antigen LFA-1. Blood. 1985 Oct;66(4):882–890. [PubMed] [Google Scholar]
  27. Sasada M., Johnston R. B., Jr Macrophage microbicidal activity. Correlation between phagocytosis-associated oxidative metabolism and the killing of Candida by macrophages. J Exp Med. 1980 Jul 1;152(1):85–98. doi: 10.1084/jem.152.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sinaceur J., Ribière C., Nordmann J., Nordmann R. Desferrioxamine: a scavenger of superoxide radicals? Biochem Pharmacol. 1984 May 15;33(10):1693–1694. doi: 10.1016/0006-2952(84)90296-x. [DOI] [PubMed] [Google Scholar]
  29. Starke P. E., Farber J. L. Ferric iron and superoxide ions are required for the killing of cultured hepatocytes by hydrogen peroxide. Evidence for the participation of hydroxyl radicals formed by an iron-catalyzed Haber-Weiss reaction. J Biol Chem. 1985 Aug 25;260(18):10099–10104. [PubMed] [Google Scholar]
  30. Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
  31. Todd R. F., 3rd, Torchia R. A., Peterson K. E., Leeman E. L. Binding and lysis of antibody-coated human erythrocytes by activated human monocytes. Clin Immunol Immunopathol. 1984 Mar;30(3):413–429. doi: 10.1016/0090-1229(84)90027-8. [DOI] [PubMed] [Google Scholar]
  32. Walker W. S., Demus A. Antibody-dependent cytolysis of chicken erythrocytes by an in vitro-established line of mouse peritoneal macrophages. J Immunol. 1975 Feb;114(2 Pt 2):765–769. [PubMed] [Google Scholar]
  33. Ward R. H., Lachmann P. J. Monoclonal antibodies which react with lymphocyte-lysed target cells and which cross-react with complement-lysed ghosts. Immunology. 1985 Sep;56(1):179–188. [PMC free article] [PubMed] [Google Scholar]
  34. Wright S. D., Meyer B. C. Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes. J Immunol. 1986 Mar 1;136(5):1759–1764. [PubMed] [Google Scholar]

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