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Interaction of mammalian cells with polymorphonuclear leukocytes: Relative sensitivity to monolayer disruption and killing

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Abstract

Monolayers of murine fibrosarcoma cells that had been treated either with histone-opsonized streptococci, histone-opsonizedCandida globerata, or lipoteichoic acid-anti-lipoteichoic acid complexes underwent disruption when incubated with human polymorphonuclear leukocytes (PMNs). Although the architecture of the monolayers was destroyed, the target cells were not killed. The destruction of the monolayers was totally inhibited by proteinase inhibitors, suggesting that the detachment of the cells from the monolayers and aggregation in suspension were induced by proteinases released from the activated PMNs. Monolayers of normal endothelial cells and fibroblasts were much more resistant to the monolayer-disrupting effects of the PMNs than were the fibrosarcoma cells. Although the fibrosarcoma cells were resistant to killing by PMNs, killing was promoted by the addition of sodium azide (a catalase inhibitor). This suggests that the failure of the PMNs to kill the target cells was due to catalase inhibition of the hydrogen peroxide produced by the activated PMNs. Target cell killing that occurred in the presence of sodium azide was reduced by the addition of a “cocktail” containing methionine, histidine, and deferoxamine mesylate, suggesting that hydroxyl radicals but not myeloperoxidase-catalayzed products were responsible for cell killing. The relative ease with which the murine fibrosarcoma cells can be released from their substratum by the action of PMNs, coupled with their insensitivity to PMN-mediated killing, may explain why the presence of large numbers of PMNs at the site of tumors produced in experimental animals by the fibrosarcoma cells is associated with an unfavorable outcome.

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References

  1. Ginsburg, I., M. N. Sela, A. Moraq, Z. Ravid, Z. Duchan, M. Ferne, S. Rabinowitz-Bergner, P. Page-Thomas, P. Davies, P. Nichols, I. Humes, andR. Bonney. 1981. The role of leukocyte factors and cationic polyelectrolytes in the phagocytosis of group A streptococci andCandida albicans by neutrophils, macrophages, fibroblasts and epithelial cells. Modulation by anionic polyelectrolytes in relation to the pathogenesis of chronic inflammation.Inflammation 5:289–312.

    Google Scholar 

  2. Ginsburg, I., N. Mor, M. Resnick, andH. Bercovier. 1986. Polycationic agents facilitate endocytosis of microorganisms by amobae.Eur. J. Cell. Biol. 41:130–133.

    Google Scholar 

  3. Ginsburg, I., R. Borinski, M. Lahav, K. E. Glllert, S. Falkenberg, M. Winkler, andS. Muller. 1982. Bacteria and zymozan opsonized with histone, dcxtran sulfate and polyanethole sulfonate trigger intense cheroilnminescence in human blood leukocytes and platelets and in mouse peritoneal macrophages. Modulation by metabolic inhibitors in relation to leukocytebacteria interactions in inflammatory sites.Inflammation 6:343–364.

    Google Scholar 

  4. Ginsburg, I., R. Borinski, D. Malamud, F. Struckmayer, andV. Fumetzek. 1985. Chemiluminescence and Superoxide generation by leukocytes stimulated by polyelectrolyte-opsonized bacteria. Role of polyarginine, polylysine, polyhistidine, cytochalasins and inflammatory exudates as modulators of the oxygen burst.Inflammation 91:245–271.

    Google Scholar 

  5. Ginsburg, I., R. Borinski, M. Lahav, Y. Matzner, I. Elliason, P. Christensen, andD. Malamud. 1984. Poly-l-arginine and an N-formylated chemotactic peptide act synergistically with lcctins and calcium ionophone to induce intense chemiluminescence and Superoxide production in human blood leukocytes. Modulation by metabolic inhibitors, sugars, and polyelectrolytes.Inflammation 8:1–26.

    Google Scholar 

  6. Ginsburg, I., R. Borinski, andM. Pabst. 1985. NADPH and “cocktails” containing polyarginine reactivate Superoxide generation in leukocytes lysed by membrane-damaging agents.Inflammation 9:341–363.

    Google Scholar 

  7. Lahav, M., andI. Ginsburg. 1977. Effect of leukocyte hydrolases on bacteria. X The role played by leukocyte factors, cationic polyelectrolytes, and by membrane-damaging agents in the lysis onStaphylococcus aureus: Relation to chronic inflammatory processes.Inflammation 2:165–177.

    Google Scholar 

  8. Ginsburg, I., M. Lahav, andP. Giesbrecht. 1982. Effect of leukocyte hydrolases on bacteria. XVI. Activation by leukocyte factors and cationic substances of antolytic wall enzymes inStaphylococcus aureus: Modulation by anionic polyelectrolytes in relation to survival of bacteria in inflammatory exudates.Inflammation 6:269–284.

    Google Scholar 

  9. Ginsburg, I., andM. Lahav. 1983. Cationic polyelectrolytes activate antolytic wall enzymes inStaphylococcus aureus: Modulation by anionic polyelectrolytes in relation to the survival of bacterial constituents in tissues.In The Target of Penicillin Action. Walter de Grayer, Berlin, 341–346.

    Google Scholar 

  10. Ginsburg, I. 1987. Cationic polyelectrolytes: A new look at their possible roles as opsonins, as stimulators of the respiratory burst in leukocytes, in bacteriolysis and as modulators of immune complex disease.Inflammation 11:489–515.

    Google Scholar 

  11. Warren, J. S., P. A. Ward, K. J. Johnson, andI. Ginsburg. 1987. Modulation of acute immune complex-mediated tissue injury by the presence of polygenic substances.Am. J. Pathol. 128:67–78.

    Google Scholar 

  12. Ginsburg, I., S. E. G. Fligiel, R. G. Kunkel, B. Riser, andJ. Varani. 1987. Phagocytosis ofCandida albicans enhances malignant behavior of murine tumor cells.Science 238:1573–1575.

    Google Scholar 

  13. Becker, E. L. 1988. The cytotoxic action of neutrophils on mammalian cells in vitro.Prog. Allergy 40:132–208.

    Google Scholar 

  14. Bevilacgua, M. P., J. S. Pober, M. E. Wheeler, R. S. Cotran, andM. A. Gimbrone, Jr. 1985. Interleukin 1 acts on cultured human vascular endothelium to increase the adhesion of polymorphonuclear leukocytes, monocytes and related cell lines.J. Clin. Invest. 76:2003–2010.

    Google Scholar 

  15. Arnaout, M. A., R. M. Hakin, R. F. Todd, N. Dana, andH. R. Colten. 1985. The granulopenia of hemodialysis is associated with increased surface expression of a graaulocyte adhesion-promoting glycoprotein (Mol).N. Engl. J. Med. 312:457–462.

    Google Scholar 

  16. Dishon, T. R. Finkel, Z. Marcus, andI. Ginsburg. 1967. Cell sensitizing products of streptococci.Immunology 13:555–564.

    Google Scholar 

  17. Lopatin, D., andR. E. Kessler. 1985. Pretreatment with lipoteichoic acid sensitizes target cells to antibody-dependent cellular cytotoxicity in the presence of anti-lipoteichoic antibodies.Infect. Immun. 48:638–643.

    Google Scholar 

  18. Varani, J., F. W. Orr, andP. A. Ward. 1979. Hydrolytic enzyme activities, migratory activity and in vivo growth and metastatic potential of recent tumor isolates.Cancer Res. 39:2376–2380.

    Google Scholar 

  19. Ryan, U. S., andL. White. 1986. Microvascular endothelium, isolation with microcarriers. Arterial, venous.J. Tissue Cult. Methods 10:9–13.

    Google Scholar 

  20. Phan, S. H., J. Varani, andD. Smith. 1985. Rat lung fibroblast collagen metabolism in bleomycin-induced pulmonary fibrosis.J. Clin. Invest. 76:241–247.

    Google Scholar 

  21. Fischer, W., H. V. Kach, andP. Haas. 1983. Improved preparation of liptoteichoic acids.Eur. J. Biochem. 154:1115–1116.

    Google Scholar 

  22. Neeman, N., andI. Ginsburg. 1971. Red cell sensitizing agent of group A streptococci. II. Immunological and immunopathological properties.Israel J. Med. Sci. 8:1807–1816.

    Google Scholar 

  23. Babior, B. M., R. S. Kipnes, andJ. T. Curnutte. 1973. Biological defense mechanisms: The production by leukocytes of Superoxide, a potential bacteriocidal agent.J. Clin. Invest. 52:741–747.

    Google Scholar 

  24. Thurman, R. G., H. G. Ley, andR. Scholz. 1972. Hepatic microsomal ethanol oxidation: Hydrogen peroxide formation and the role of catalase.Eur. J. Biochem. 25:420–427.

    Google Scholar 

  25. Anson, M. L. 1940. The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin.J. Gen. Physiol. 23:79–86.

    Google Scholar 

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

    Google Scholar 

  27. Ginsburg, I., S. E. G. Fligiel, P. A. Ward, andJ. Varani. 1988. Lipoteichoic acid antilipoteichoic acid complexs induce Superoxide generation by human neutrophils.Inflammation 12:525–548.

    Google Scholar 

  28. Varani, J., S. E. G. Fligiel, G. O. Till, R. G. Kunkel, U. S. Ryan, andP. A. Ward. 1985. Pulmonary endothelial cell killing by human neutrophils. Possible involvement of hydroxyl radical.Lab. Invest. 53:656–663.

    Google Scholar 

  29. Gannon, D. E., J. Varani, S. H. Phan, J. H. Ward, J. Kaplan, G. O. Till, R. H. Simon, U. S. Ryan, andP. A. Ward. 1987. Source of iron in neutrophil-mediated killing of endothelial cells.Lab. Invest. 57:37–44.

    Google Scholar 

  30. Varani, J., M. J. Bendelow, D. E. Sealey, S. L. Kunkel, D. E. Gannon, U. S. Ryan, andP. A. Ward. 1988. Tumor necrosis factor enhances susceptibility of vascular enclothelial cells to neutrophil-mediated killing,lab. Invest. 59:292–295.

    Google Scholar 

  31. Sacks, T., C. F. Moldow, P. R. Craddock, T. K. Bowers, andH. S. Jacob. 1978. Oxygen radicals mediate endothelial cell damage by complement-stimulated granulocytes. An in vitro model of immune vascular damage.J. Clin. Invest. 61:1161–1167.

    Google Scholar 

  32. Weiss, S. J., J. Young, A. F. LoBuglio, A. Slivka, N. F. Njmeh. 1981. Role of hydrogen peroxide in neutrophil-mediated destruction of cultured endothelial cells.J. Clin. Invest. 68:714–720.

    Google Scholar 

  33. Martin, W. J. 1984. Neutrophils kill pulmonary endothelial cells by a hydrogen-peroxide-dependent pathway. An in vitro model of neutrophil-mediated lung injury.Am. Rev. Respir. Dis. 130:209–213.

    Google Scholar 

  34. Starke, P. E., andJ. L. Farber. 1985. 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. 260:10099–10106.

    Google Scholar 

  35. Fligiel, S. E. G., P. A. Ward, K. J. Johnson, andG. O. Till. 1984. Evidence for a role of hydroxyl radical in immune complex-induced vasculitis.Am. J. Pathol. 115:397–405.

    Google Scholar 

  36. Ward, P. A., G. O. Till, R. Kunkel, andC. Beauchamp. 1983. Evidence for a role of hydroxyl radical in complement and neutrophil-dependent tissue injury.J. Clin. Invest. 72:789–795.

    Google Scholar 

  37. Clarke, R. A., andS. J. Klebanoff. 1975. Neutrophil-mediated tumor cell toxicity. Role of the peroxidase system.J. Exp. Med. 141:1442–1447.

    Google Scholar 

  38. Smedley, L. A., M. G. Tonnesen, R. A. Sandhous, C. Haslett, L. A. Guthrie, R. B. Johnston, P. M. Henson, andG. S. Worthen. 1986. Neutrophil-mediated injury to endothelial cells. Enhancement by endotoxin and essential role for neutrophil elastase.J. Clin. Invest. 77:1233–1239.

    Google Scholar 

  39. Harlan, J. M., B. R. Schwartz, M. A. Reidy, S. M. Schwartz, H. D. Ochs, andL. A. Harker. 1985. Activated neutrophils disrupt endothelial monolayer integrity by an oxygen radical-independent mechanism.Lab. Invest. 52:141–150.

    Google Scholar 

  40. Senior, R. M., H. Tegner, C. Kuhn, K. Ohlsson, B. C. Stascher, andJ. A. Pierce. 1977. The induction of pulmonary emphysema with human leukocyte elastase.Am. Rev. Respir. Dis. 116:465–475.

    Google Scholar 

  41. Taubman, S. B., andR. B. Cogen. 1975. Cell-detaching activity mediated by an enzyme(s) obtained from human leukocyte granules.Lab. Invest. 32:555–560.

    Google Scholar 

  42. Giraldi, T. C., C. Nisi, andG. Sava. 1977. Lysomal enzyme inhibitors and antimetastatic activity in the mouse.Eur. J. Cancer 13:1321–1323.

    Google Scholar 

  43. Petterson, J. 1977. Fibrinolysis and anti-fibrinolytic agents in the growth and spread of tumors.Cancer Treat. Rev. 4:213–217.

    Google Scholar 

  44. Alitalo, K., J. Keski-Oja, K. Hedman, andA. Vaheri. 1982. Loss of different pericellular matrix components of rat cells transformed with a T-class ts mutant of Rous sarcoma virus.Virology 119:347–357.

    Google Scholar 

  45. Vaheri, A., andD. F. Mosher. 1978. High molecular weight, cell surface-associated glycoprotein (Fibronectin) lost in malignant transformation.Biochim. Biophys. Acta 516:1–9.

    Google Scholar 

  46. Yamada, K. M., K. Oolden, andI. Pastan. 1978. Transformation-sensitive cell surface protein. Isolation, characterization and role in cellular morphology and adhesion. N.Y. Acad. Sci.312:256–267.

    Google Scholar 

  47. Simon, R. H., C. H. Scoggin, andD. Peterson. 1981. Hydrogen peroxide causes the fatal injury to human flbroblasts exposed to oxygen radicals.J. Biol. Chem. 256:7181–7186.

    Google Scholar 

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Authors and Affiliations

  1. Department of Oral Biology, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel

    Isaac Ginsburg

  2. Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan

    Douglas F. Gibbs & James Varani

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  1. Isaac Ginsburg
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Additional information

Supported by a research grant from Dr. S.M. Robbins, Cleveland, Ohio; by research grant IM-432 from the American Cancer Society and by grants HL-28872, HL-37963 and GM-29501 from the National Institutes of Health, Bethesda, MD. Isaac Ginsburg was a visiting professor at the Department of Pathology in 1987–1988.

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Ginsburg, I., Gibbs, D.F. & Varani, J. Interaction of mammalian cells with polymorphonuclear leukocytes: Relative sensitivity to monolayer disruption and killing. Inflammation 13, 529–542 (1989). https://doi.org/10.1007/BF00916759

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