Analysis of induced sputum after air and ozone exposures in healthy subjects
Abstract
Exposure of healthy subjects to ozone is associated with increases in cellular and biochemical markers of inflammation in bronchoalveolar lavage fluid. To determine if analysis of induced sputum might similarly reveal the pulmonary inflammatory effects of ozone exposure, we performed cellular and biochemical analysis of induced sputum collected 4 hr after air and ozone (0.4 ppm for 2 hr) exposures from 10 healthy subjects (age 30.0 +/- 5.0 years; 5 females) in a randomized crossover study in which exposures were separated by 2 weeks. We found that the total number of nonsquamous cells was significantly higher after ozone exposure than after air exposure (7.4 vs 3.9 x 10(5)/ml, P < 0.05) as was the percentage of the nonsquamous cells that were neutrophils (80.0 +/- 7.0% vs 51.0 +/- 20.0%, P < 0.05) and the levels of myeloperoxidase in the sputum fluid phase (1.6 +/- 0.6 vs 1.3 +/- 0.6 microg/ml, P < 0.05). In addition, IL-6 and IL-8 levels were higher after ozone than after air exposures, but not significantly so (44.5 +/- 32.4 pg/ml vs 26.8 +/- 30.7 pg/ml, P = 0.11; 1.5 +/- 0.5 ng/ml vs 1.1 +/- 0.6 ng/ml, P = 0.09). Mucin-like glycoprotein levels were also not significantly different between exposures (1.6 +/- 0.9 mg/ml vs 1.3 +/- 1.0 mg/ml, P = 0.26). We conclude that analysis of induced sputum is a useful noninvasive method for studying the pulmonary response to ozone exposure in healthy subjects.
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Ozone exposure resulted in significant neutrophilic inflammation, reflected with increased levels of myeloperoxidase in the supernatant of induced sputum samples from healthy subjects.
“Ozone exposure resulted in significant neutrophilic inflammation, reflected with increased levels of myeloperoxidase (MPO) in the supernatant of induced sputum samples from healthy subjects (39, 40).”
From Ozone pollution: A Major health hazard worldwide (2019) · cites this paper as [39]
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Ozone pollution: A Major health hazard worldwide
Zhang, J. (., Wei, Y., & Fang, Z. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.02518 · Cited by 828 · see citing works
“Ozone exposure resulted in significant neutrophilic inflammation, reflected with increased levels of myeloperoxidase (MPO) in the supernatant of induced sputum samples from healthy subjects (39, 40).”
BackgroundSzefler, S. J., Wenzel, S. E., Brown, R. H., ERZURUM, S. C., Fahy, J. V., Hamilton, R. G., Hunt, J. F., Kita, H., Liu, A. H., Panettieri, R. A., et al. (2012) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2011.12.979 · Cited by 395 · see citing works
Self-citationThe role of eosinophils and neutrophils in inflammation
Sampson, A. P. (2000) · Clinical & Experimental Allergy · doi:10.1046/j.1365-2222.2000.00092.x · Cited by 259 · see citing works
Gong, H., Linn, W. S., Sioutas, C., Terrell, S. L., Clark, K. W., Anderson, K. R., & Terrell, L. L. (2003) · Inhalation Toxicology · doi:10.1080/08958370304455 · Cited by 165 · see citing works
“Immediately following the day-2 testing sequence, the subject underwent sputum induction (Fahy et al., 1996).”
MethodNordenhäll, C., Pourazar, J., Blomberg, A., Levin, J.-O., Sandström, T., & Adelroth, E. C. (2000) · European Respiratory Journal · doi:10.1034/j.1399-3003.2000.01512.x · Cited by 156 · see citing works
“In air pollution research, induced sputum has previously proved valuable in the evaluation of the airway effects of ozone [ 26 ] and nitrogen dioxide [27].”
BackgroundNoninvasive methods for assessment of airway inflammation in occupational settings
Quirce, S., Lemière, C., Frederic de, B. L. A. Y., del Pozo, V., van Wijk, R. G., Maestrelli, P., Pauli, G., Pignatti, P., Raulf‐Heimsoth, M., Sastre, J., et al. (2009) · Allergy · doi:10.1111/j.1398-9995.2009.02274.x · Cited by 131 · see citing works
“Exposure to isocyanates (5), acrylates or flour (6) does not seem to induce eosinophilic or neutrophilic inflammation in healthy subjects, but a neutrophilic inflammatory response has been found in the induced sputum of healthy subjects after short-term exposure to irritant agents such as ozone (57), diesel exhaust (5…”
BackgroundNightingale, J. A., Rogers, D. F., & Barnes, P. J. (1999) · Thorax · doi:10.1136/thx.54.12.1061 · Cited by 119 · see citing works
“accompanied by a neutrophilic inflammatory response”
Ozone-induced Airway Inflammatory Changes Differ between Individuals and Are Reproducible
Holz, O., Jörres, R. A., TIMM, P., et al. (1999) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/ajrccm.159.3.9806098 · Cited by 113 · see citing works
“Fahy and colleagues (7) studied 10 healthy subjects who were exposed to similar conditions [400 ppb, 2 h, 25 L/ (min ? m(2))], with sputum induction 4 h later.”
BackgroundMethodResultInfluentialGong, H., Linn, W. S., Clark, K. W., et al. (2008) · Inhalation Toxicology · doi:10.1080/08958370801911340 · Cited by 108 · see citing works
“Sputum induction was performed on day 2 only, after completion of other testing and prophylactic administration of a bronchodilator, by the methods of Fahy et al. (1995).”
MethodOzone Exposure Increases Eosinophilic Airway Response Induced by Previous Allergen Challenge
Vagaggini, B., Taccola, M., Cianchetti, S., et al. (2002) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/rccm.2201013 · Cited by 104 · see citing works
Analysis of fluidphase mediators
Leader of the Working Group: M.M. Kelly1, Members of the Working Group:, Keatings, V. M., et al. (2002) · European Respiratory Journal · doi:10.1183/09031936.02.00002402 · Cited by 101 · see citing works
Induced sputum, eosinophilic bronchitis, and chronic obstructive pulmonary disease.
Hargreave, F., & Leigh, R. (1999) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/ajrccm.160.supplement_1.14 · Cited by 103 · see citing works
Gong, H., Linn, W. S., Terrell, S. L., Anderson, K. R., Clark, K. W., Sioutas, C., Cascio, W. E., Alexis, N. E., & Devlin, R. B. (2004) · Inhalation Toxicology · doi:10.1080/08958370490499906 · Cited by 96 · see citing works
“After the day-2 testing sequence, sputum was induced with 3% saline aerosol inhalation and assayed for cellular and biochemical markers of airway inflammation (Fahy et al., 1996).”
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“A sputum neutrophilia also occurs in smokers with (10,21) or without (5) chronic airflow limitation, with viral infections (22), and with exposure to endotoxin (23) or ozone (24).”
BackgroundInduced sputum cell counts: their usefulness in clinical practice
Jayaram, L., Parameswaran, K., Sears, M. R., et al. (2000) · European Respiratory Journal · doi:10.1034/j.1399-3003.2000.16a27.x · Cited by 89 · see citing works
Gong, H., Linn, W. S., Clark, K. W., et al. (2005) · Inhalation Toxicology · doi:10.1080/08958370590904481 · Cited by 83 · see citing works
“During the day 2 visit, spirometry, sputum induction (Fahy et al., 1996) with 3% saline, and total sputum cell counts were performed.”
MethodOzone enhances the airway inflammation initiated by diesel exhaust
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No Effect of Inhaled Budesonide on the Response to Inhaled Ozone in Normal Subjects
Nightingale, J. A., Rogers, D. F., Chung, K. F., & Barnes, P. J. (2000) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/ajrccm.161.2.9905031 · Cited by 73 · see citing works
“A previous study using induced sputum to study responses to 400 ppb ozone showed significant increases in MPO and nonsignificant increases in IL-8 ( 7 ), as seen herein.”
BackgroundResultInfluentialRespiratory Responses to Ozone Exposure. MOSES (The Multicenter Ozone Study in Older Subjects)
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“Several controlled human exposure studies have documented that inhalation of high ambient levels of ozone results in airway neutrophilia (2, 3, 32–34), which is detectable in induced sputum (35, 36).”
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“Nitrogen dioxide (NO 2 ) and ozone seem to be involved in this process [15, 16 ].”
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Parameswaran, K., Pizzichini, E., Pizzichini, M. M. M., et al. (2000) · European Respiratory Journal · doi:10.1034/j.1399-3003.2000.15.10.x · Cited by 67 · see citing works
New insights in neutrophilic asthma
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Ozone-induced inflammation assessed in sputum and bronchial lavage fluid from asthmatics:
Hiltermann, T. J. N., Lapperre, T. S., van Bree, L., et al. (1999) · Free Radical Biology and Medicine · doi:10.1016/s0891-5849(99)00191-4 · Cited by 57 · see citing works
“Recently, new techniques such as sputum induction have been developed that may be more appropriate to study ozone-induced lower airways inflammation than nasal lavage [1,13].”
MethodBudesonide Reduces Neutrophilic but Not Functional Airway Response to Ozone in Mild Asthmatics
Vagaggini, B., Taccola, M., Conti, I., et al. (2001) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/ajrccm.164.12.2009090 · Cited by 49 · see citing works
Gong, H., Linn, W. S., Terrell, S. L., et al. (2001) · CHEST Journal · doi:10.1378/chest.119.2.402 · Cited by 48 · see citing works
Airway inflammatory response to ozone in subjects with different asthma severity
Vagaggini, B., Carnevali, S., Macchioni, P., et al. (1999) · European Respiratory Journal · doi:10.1034/j.1399-3003.1999.13b09.x · Cited by 45 · see citing works
“FAHY et al. [ 21 ] showed that exposure of healthy subjects to 0.4 ppm O3 for 2 h was associated with significantly greater increases in sputum neutrophil percentage and in myeloperoxidase (MPO) concentration than air exposure.”
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Induced Sputum for the Investigation of Airway Inflammation: Evidence for Its Clinical Application
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“Neutrophilic inflammation occurs in cigarette smokers (9,26), during bacterial and viral infections (27), and after the inhalation of endotoxin (28) and some pollutants such as ozone (29).”
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“In a polluted environment (primarily found in urban areas), ozone can reach unhealthy levels during sunny days, even though high levels of ozone have been observed during cold days (Calfapietra et al. 2009, 2013; Chang et al. 2010; Fahy et al. 1995).”
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“Confirming the work of FAHY et al. [ 6 ] and HILTERMANN et al. [7], the current authors demonstrated in both conditions a pre- to post-exposure increase in per cent neutrophils in induced sputum.”
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- Authors
- Fahy, J. V., Wong, H. H., Liu, J. T., & Boushey, H. A.
- Affiliations
- University of California, San Francisco (US)
- Venue
- Environmental Research
- Publisher
- Elsevier BV
- Published
- 1995-08
- Volume
- 70
- Issue
- 2
- Pages
- 77-83
- ISSN
- 0013-9351 (ISSN-L)
- OpenAlex
- W2090582545
- Cited by
- 54
- References
- 0
- Citations by year
- 2026: 1 · 2024: 3 · 2019: 2 · 2018: 1 · 2017: 2 · 2015: 1 · 2012: 1
- FWCI
- 0.40
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Environmental Research (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Air Quality and Health Impacts
- Health Sciences › Medicine › Physiology › Asthma and respiratory diseases
- Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Indoor Air Quality and Microbial Exposure
- Keywords
- ozone exposure, induced sputum, air pollution, lung inflammation, neutrophils, myeloperoxidase, interleukin-6, interleukin-8, mucin-like glycoprotein, healthy volunteers, non-invasive biomarkers
- MeSH terms
- Adult, Airway Resistance, Cell Count, Eosinophils, Epithelial Cells, Epithelium, Female, Humans, Lymphocytes, Macrophages, Male, Mucins, Peroxidase, Neutrophils, Ozone, Surveys and Questionnaires, Spirometry, Sputum, Interleukin-6, Single-Blind Method, Interleukin-8, Cross-Over Studies
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