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

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.

[1]  K. Driscoll.  ROLE OF INFLAMMATION IN THE DEVELOPMENT OF RAT LUNG TUMORS IN RESPONSE TO CHRONIC PARTICLE EXPOSURE , 1996 .

[2]  H. Koren,et al.  Biomarkers of inflammation in ozone-exposed humans. Comparison of the nasal and bronchoalveolar lavage. , 1990, The American review of respiratory disease.

[3]  H. van Loveren,et al.  Nasal lavage biomarkers in air pollution epidemiology. , 1997, Archives of toxicology. Supplement. = Archiv fur Toxikologie. Supplement.

[4]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[5]  H. Gallati,et al.  [Horseradish peroxidase: kinetic studies and optimization of peroxidase activity determination using the substrates H2O2 and 3,3',5,5'-tetramethylbenzidine]. , 1985, Journal of clinical chemistry and clinical biochemistry. Zeitschrift fur klinische Chemie und klinische Biochemie.

[6]  K. Kohn,et al.  Activated neutrophils induce prolonged DNA damage in neighboring cells. , 1989, Carcinogenesis.

[7]  E. Pick,et al.  A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture. , 1980, Journal of immunological methods.

[8]  F. Seiler,et al.  Formation and persistence of 8-oxoguanine in rat lung cells as an important determinant for tumor formation following particle exposure. , 1997, Environmental health perspectives.

[9]  H Greim,et al.  Toxicity of fibers and particles. Report of the workshop held in Munich, Germany, 26-27 October 2000. , 2001, Inhalation toxicology.

[10]  J. Doroshow,et al.  Modification of DNA bases in chromatin of intact target human cells by activated human polymorphonuclear leukocytes. , 1993, Cancer research.

[11]  J. Mauderly,et al.  Toxic and carcinogenic effects of solid particles in the respiratory tract , 1994 .

[12]  P. Borm,et al.  Neutrophils cause oxidative DNA damage in alveolar epithelial cells. , 1999, Free radical biology & medicine.

[13]  A. Kettle,et al.  Inside the neutrophil phagosome: oxidants, myeloperoxidase, and bacterial killing. , 1998, Blood.

[14]  P. Borm,et al.  Ambient Particulate Matter Induces Oxidative Dna Damage in Lung Epithelial Cells , 2000, Inhalation toxicology.

[15]  H. Gallati,et al.  Peroxidase aus Meerrettich: Kinetische Studien und Optimierung der Peroxidase-Aktivitätsbestimmung mit den Substraten H2C2 und 3,3',5,5'-Tetramethylbenzidin , 1985 .

[16]  R. Santella,et al.  8-hydroxy-2'-deoxyguanosine, a major mutagenic oxidative DNA lesion, and DNA strand breaks in nasal respiratory epithelium of children exposed to urban pollution. , 1999, Environmental health perspectives.

[17]  G. Lubec,et al.  Aromatic hydroxylation in nasal lavage fluid following ambient ozone exposure. , 1997, Free radical biology & medicine.

[18]  H. Ochi,et al.  Quantitative immunohistochemical determination of 8-hydroxy-2'-deoxyguanosine by a monoclonal antibody N45.1: its application to ferric nitrilotriacetate-induced renal carcinogenesis model. , 1997, Laboratory investigation; a journal of technical methods and pathology.

[19]  P. Borm,et al.  Oxidative DNA damage in peripheral blood lymphocytes of coal workers , 1995, International archives of occupational and environmental health.

[20]  J. Douwes,et al.  Longitudinal changes in inflammatory markers in nasal lavage of cotton workers , 1998, International archives of occupational and environmental health.

[21]  S. Weitzman,et al.  Inflammation and cancer: role of phagocyte-generated oxidants in carcinogenesis. , 1990, Blood.

[22]  B. Babior.  Phagocytes and oxidative stress. , 2000, The American journal of medicine.

[23]  H. Rosen,et al.  Antimicrobial activity of myeloperoxidase. , 1984, Methods in enzymology.

[24]  B. Halliwell,et al.  Nitrite-induced deamination and hypochlorite-induced oxidation of DNA in intact human respiratory tract epithelial cells. , 2000, Free radical biology & medicine.

[25]  S. Weiss.  Tissue destruction by neutrophils. , 1989, The New England journal of medicine.

[26]  S. Latt,et al.  Stimulated human phagocytes produce cytogenetic changes in cultured mammalian cells. , 1983, The New England journal of medicine.

[27]  S. Weitzman,et al.  Phagocytes as carcinogens: malignant transformation produced by human neutrophils. , 1985, Science.

[28]  P. Borm,et al.  Nasal inflammatory and respiratory parameters in human volunteers during and after repeated exposure to chlorine. , 2000, The European respiratory journal.

[29]  M. O’Reilly,et al.  Pulmonary chemokine and mutagenic responses in rats after subchronic inhalation of amorphous and crystalline silica. , 2000, Toxicological sciences : an official journal of the Society of Toxicology.

[30]  S. Linn,et al.  Formation, Prevention, and Repair of DNA Damage by Iron/Hydrogen Peroxide* , 1997, The Journal of Biological Chemistry.

[31]  J. Jackson,et al.  Oxidant-induced DNA damage of target cells. , 1988, The Journal of clinical investigation.

[32]  R E Durand,et al.  Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay. , 1990, Radiation research.

[33]  H. Rosen,et al.  [52] Antimicrobial activity of myeloperoxidase , 1984 .

[34]  R. Simon,et al.  Neutrophil-induced injury of rat pulmonary alveolar epithelial cells. , 1986, The Journal of clinical investigation.

[35]  P. Steerenberg,et al.  Nasal lavage as tool for health effect assessment of photochemical air pollution , 1996, Human & experimental toxicology.

[36]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[37]  J L Mauderly,et al.  Comparative Pulmonary Toxicities and Carcinogenicities of Chronically Inhaled Diesel Exhaust and Carban Black in F344 Rats , 1995, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[38]  J. Carter,et al.  Effects of particle exposure and particle-elicited inflammatory cells on mutation in rat alveolar epithelial cells. , 1997, Carcinogenesis.

[39]  P. Borm,et al.  Neutrophils amplify the formation of DNA adducts by benzo[a]pyrene in lung target cells. , 1997, Environmental health perspectives.

[40]  B. Halliwell,et al.  Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. , 1996, The Biochemical journal.

[41]  T. Standiford,et al.  Interleukin-8 (IL-8): the major neutrophil chemotactic factor in the lung. , 1991, Experimental lung research.