Molecular and Cellular Biochemistry

, Volume 234, Issue 1, pp 143–151

Mechanisms of neutrophil-induced DNA damage in respiratory tract epithelial cells

  • Ad M. Knaapen
  • Roel P.F. Schins
  • Dünya Polat
  • Andrea Becker
  • Paul J.A. Borm
Article

Abstract

Reactive oxygen species (ROS) released by neutrophils have been suggested to play an important role in cancer development. Since the mechanisms underlying this effect in the respiratory tract are still unclear, we evaluated DNA damage induced by neutrophils in respiratory tract epithelial cells in vitro and in vivo. For in vitro studies, rat lung epithelial cells (RLE) were co-incubated with activated neutrophils, neutrophil-conditioned medium, or hydrogen peroxide. For in vivo studies, we considered the human nose as a target organ, comparing neutrophilic inflammation in the nasal lavage fluid with the oxidative DNA lesion 8-hydroxydeoxyguanosine (8-OHdG) in epithelial cells obtained by nasal brush. Our in vitro data show that human neutrophils are able to induce both 8-OHdG and strand breaks in DNA from RLE cells. Our data also suggest that DNA damage induced by neutrophils is inhibited when neutrophil-derived H2O2 is consumed by myeloperoxidase. In contrast, in the nose no association between neutrophil numbers and 8-OHdG was found. Therefore, it remains unclear whether neutrophils pose a direct genotoxic risk for the respiratory tract epithelium during inflammation, and more in vivo studies are needed to elucidate the possible association between neutrophils and genotoxicity in the lung.

DNA damage genotoxicity 8-hydroxydeoxyguanosine inflammation nasal brush nasal lavage neutrophils reactive oxygen species respiratory tract epithelial cells strand breaks 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Weitzman SA, Gordon LI: Inflammation and cancer: Role of phagocyte-generated oxidants in carcinogenesis. Blood 76: 655–663, 1990PubMedGoogle Scholar
  2. 2.
    Dungworth DL, Mohr U, Heinrich U, Ernst H, Kittel B: Pathologic effects of inhaled particles in rat lungs: Associations between inflammatory and neoplastic processes. In: U. Mohr (ed). Toxic and Carcinogenic Effects of Solid Particles in the Respiratory Tract. ILSI press, Washington, 1994, pp 75–98Google Scholar
  3. 3.
    Driscoll KE, Deyo LC, Carter JM, Howard BW, Hassenbein DG, Bertram TA: Effects of particle exposure and particle-elicited inflammatory cells on mutation in rat alveolar epithelial cells. Carcinogenesis 18: 423–430, 1997PubMedGoogle Scholar
  4. 4.
    Babior BM: Phagocytes and oxidative stress. Am J Med 109: 33–44, 2000CrossRefPubMedGoogle Scholar
  5. 5.
    Weitberg AB, Weitzman SA, Destrempes M, Latt SA, Stossel TP: Stimulated human phagocytes produce cytogenetic changes in cultured mammalian cells. New Eng J Med 308: 26–29, 1983PubMedGoogle Scholar
  6. 6.
    Weitzman SA, Weitberg AB, Clark EP, Stossel TP: Phagocytes as carcinogens: Malignant transformation produced by human neutrophils. Science 227: 1231–1233, 1984Google Scholar
  7. 7.
    Wiseman H, Halliwell B: Damage to DNA by reactive oxygen and nitrogen species: Role in inflammatory disease and progression to cancer. Biochem J 313: 17–29, 1996PubMedGoogle Scholar
  8. 8.
    Shacter E, Beecham EJ, Covey JM, Kohn KW, Potter M: Activated neutrophils induce prolonged DNA damage in neighboring cells. Carcinogenesis 9: 2297–2304, 1988PubMedGoogle Scholar
  9. 9.
    Schraufstätter I, Hyslop PA, Jackson JH, Cochrane CG: Oxidant-induced DNA damage of target cells. J Clin Invest 82: 1040–1050, 1988PubMedGoogle Scholar
  10. 10.
    Dizdaroglu M, Olinski R, Doroshow JH, Akman SA: Modification of DNA bases in chromatin of intact target human cells by activated human polymorphonuclear leukocytes. Cancer Res 53: 1296–1272, 1993Google Scholar
  11. 11.
    Knaapen AM, Seiler F, Schilderman PAEL, Nehls P, Bruch J, Schins RPF, Borm PJA: Neutrophils cause oxidative DNA damage in alveolar epithelial cells. Free Rad Biol Med 27: 234–240, 1999CrossRefPubMedGoogle Scholar
  12. 12.
    Nehls P, Seiler F, Rehn B, Greferath R, Bruch J: Formation and persistence of 8-oxoguanine in rat lung cells as an important determinant for tumor formation following particle exposure. Environ Health Perspect 105(suppl 5): 1291–1296, 1997PubMedGoogle Scholar
  13. 13.
    Driscoll KE: Role of inflammation in the development of rat lung tumors in response to chronic particle exposure. Inhal Toxicol 8(suppl): 139–153, 1996Google Scholar
  14. 14.
    Johnston CJ, Driscoll KE, Finkelstein JN, Baggs R, O'Reilly MA, Carter J, Gelein R, Oberdörster G: Pulmonary chemokine and mutagenic responses in rats after subchronic inhalation of amorphous and crystalline silica. Toxicol Sci 56: 405–413, 2000CrossRefPubMedGoogle Scholar
  15. 15.
    Greim H, Borm PJA, Schins RPF, Donaldson K, Driscoll KE, Hartwig A, Kuempel E, Oberdörster G, Speit G: Toxicity of fibers and particles. Report of the workshop held in Munich, Germany, 26- 27th October 2000. Inhal Toxicol 13: 101–119, 2001Google Scholar
  16. 16.
    Driscoll KE, Carter JM, Iype PT, Kumari HL, Crosby LL, Aardema MJ, Isfort RJ, Cody D, Chestnut MH, Burns JL, Le Boeuf RA: Establishment of immortalized alveolar type II cell lines from adult rat lungs. In Vitro Cell Dev Biol 31: 516–527, 1995Google Scholar
  17. 17.
    Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Meth 65: 517–527, 1983Google Scholar
  18. 18.
    Borm PJA, Knaapen AM, Schins RPF, Godschalk RWL, Van Schooten FJ: Neutrophils amplify the formation of DNA-adducts by benzo-[a]pyrene in lung target cells. Environ Health Perspect 105(suppl 5): 1089–1093, 1997PubMedGoogle Scholar
  19. 19.
    Pick E, Keisari YA: simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture. J Immunol Meth 38: 161–170, 1980Google Scholar
  20. 20.
    Gallati H, Pracht I: Horseradish peroxidase: Kinetic studies and optimization of peroxidase activity determination using substrates H2O2 and 3,3′,5,5′-tetramethylbenzidine. J Clin Chem Clin Biochem 23: 453–460, 1985PubMedGoogle Scholar
  21. 21.
    Klebanoff SJ, Waltersdorph AM, Rosen H: Antimicrobial activity of myeloperoxidase. Meth Enzymol 105: 399–403, 1984PubMedGoogle Scholar
  22. 22.
    Singh NP, McCoy MT, Tice RR, Schneider ELA: Simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175: 184–191, 1988PubMedGoogle Scholar
  23. 23.
    Olive PL, Banath JP, Durand RE: Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the 'comet' assay. Radiat Res 122: 86–94, 1990PubMedGoogle Scholar
  24. 24.
    Keman S, Jetten M, Douwes J, Borm PJA: Longitudinal changes of inflammatory markers in nasal lavage of cotton workers. Int Arch Occup Environ Health 71: 131–137, 1998PubMedGoogle Scholar
  25. 25.
    Schins RPF, Schilderman PAEL, Borm PJA: Oxidative DNA damage in peripheral blood lymphocytes of coal workers. Int Arch Occup Environ Health 67: 153–157, 1995PubMedGoogle Scholar
  26. 26.
    Knaapen AM, Schins RPF, Steinfartz Y, Höhr D, Dunemann L, Borm PJA: Ambient particulate matter induces oxidative DNA damage in lung epithelial cells. Inhal Toxicol 12(suppl 3): 125–132, 2000Google Scholar
  27. 27.
    Toyokuni S, Tanaka T, Hattori Y, Nishiyama Y, Yoshida A, Uchida K, Hiai H, Ochi H, Osawa T: Quantitative immunohistochemical determination of 8-hydroxy-2′-deoxyguanosine by a monoclonal antibody N45.1: Its application to ferric nitrolotriacetate-induced renal carcinogenesis model. Lab Invest 76: 365–374, 1997PubMedGoogle Scholar
  28. 28.
    Nikula KJ, Snipes MB, Barr EB, Griffith WC, Henderson RF, Mauderly JL: Comparative pulmonary toxicities and carcinogenicities of chronically inhaled diesel exhaust and carbon black in F344 rats. Fundam Appl Toxicol 25: 80–94, 1995PubMedGoogle Scholar
  29. 29.
    Henle ES, Linn S: Formation, prevention, and repair of DNA damage by iron/hydrogen peroxide. J Biol Chem 272: 19095–19098, 1997CrossRefPubMedGoogle Scholar
  30. 30.
    Hampton MB, Kettle AJ, Winterbourne CC: Inside the neutrophil phagosome: Oxidants, myeloperoxidase, and bacterial killing. Blood 92: 3007–3017, 1998PubMedGoogle Scholar
  31. 31.
    Spencer JPE, Whiteman M, Jenner A, Halliwell B: Nitrite-induced deamination and hypochlorite-induced oxidation of DNA in intact human respiratory tract epithelial cells. Free Rad Biol Med 28: 1039–1050, 2000PubMedGoogle Scholar
  32. 32.
    Weiss SJ: Tissue destruction by neutrophils. New Eng J Med 320: 365–376, 1989PubMedGoogle Scholar
  33. 33.
    Schins RPF, Emmen H, Hoogendijk L, Borm PJA: Nasal inflammatory and respiratory parameters in human volunteers during and after repeated exposure to chlorine. Eur Respir J 16: 626–632, 2000CrossRefPubMedGoogle Scholar
  34. 34.
    Calderó n-Garciduenas L, Wen-Wang L, Zhang YJ, Rodriguez-Alcaraz A, Osnaya N, Villareal-Calderó n A, Santella RM: 8-Hydroxy-2′-deoxyguanosine, a major mutagenic oxidative DNA lesion, and DNA strand breaks in nasal respiratory epithelium of children exposed to urban pollution. Environ Health Perspect 107: 469–474, 1999Google Scholar
  35. 35.
    Graham DE, Koren HS: Biomarkers of inflammation in ozone-exposed humans. Comparison of the nasal and bronchoalveolar lavage. Am Rev Respir Dis 142: 152–156, 1990PubMedGoogle Scholar
  36. 36.
    Kunkel SL, Standiford T, Kasahara K, Strieter RM: Interleukin-8 (IL-8), the major neutrophil chemotactic factor in the lung. Exp Lung Res 17: 17–23, 1991PubMedGoogle Scholar
  37. 37.
    Steerenberg PA, Fischer PH, van Bree L, van Loveren H: Nasal lavage biomarkers in air pollution epidemiology. Arch Toxicol 19(suppl): 207–216, 1997Google Scholar
  38. 38.
    Steerenberg PA, Fischer PH, Gmelig Meyling F, Willighagen J, Geerse E, van de Vliet H, Ameling C, Boink AB, Dormans JA, van Bree L, van Loveren H: Nasal lavage as tool for health effect assessment of photochemical air pollution. Hum Exp Toxicol 15: 111–119, 1996PubMedGoogle Scholar
  39. 39.
    Simon RH. DeHart PD, Todd III RF: Neutrophil-induced injury of rat pulmonary alveolar epithelial cells. J Clin Invest 78: 1375–1386, 1986PubMedGoogle Scholar
  40. 40.
    Frischer T, Pullwitt A, Kuhr J, Meinert R, Haschke N, Studnicka M, Lubec G: Aromatic hydroxylation in nasal lavage fluid following ambient ozone exposure. Free Rad Biol Med 22: 201–207, 1997PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Ad M. Knaapen
    • 1
  • Roel P.F. Schins
    • 1
  • Dünya Polat
    • 1
  • Andrea Becker
    • 1
  • Paul J.A. Borm
    • 1
  1. 1.Particle Research CoreInstitute for Environmental Health (IUF)DüsseldorfGermany

Personalised recommendations